Advances in Malaria Diagnostic Methods in Resource-Limited Settings: A Systematic Review
疟疾诊断方法在资源有限地区的进展：系统性综述

Abstract  摘要

Malaria continues to pose a health challenge globally, and its elimination has remained a major topic of public health discussions. A key factor in eliminating malaria is the early and accurate detection of the parasite, especially in asymptomatic individuals, and so the importance of enhanced diagnostic methods cannot be overemphasized. This paper reviewed the advances in malaria diagnostic tools and detection methods over recent years. The use of these advanced diagnostics in lower and lower-middle-income countries as compared to advanced economies has been highlighted. Scientific databases such as Google Scholar, PUBMED, and Multidisciplinary Digital Publishing Institute (MDPI), among others, were reviewed. The findings suggest important advancements in malaria detection, ranging from the use of rapid diagnostic tests (RDTs) and molecular-based technologies to advanced non-invasive detection methods and computerized technologies. Molecular tests, RDTs, and computerized tests were also seen to be in use in resource-limited settings. In all, only twenty-one out of a total of eighty (26%) low and lower-middle-income countries showed evidence of the use of modern malaria diagnostic methods. It is imperative for governments and other agencies to direct efforts toward malaria research to upscale progress towards malaria elimination globally, especially in endemic regions, which usually happen to be resource-limited regions.
疟疾持续对全球健康构成挑战，其消除问题始终是公共卫生讨论的核心议题。实现疟疾消除的关键在于对寄生虫（尤其是无症状感染者）的早期精准检测，因此改进诊断方法的重要性再怎么强调都不为过。本文综述了近年来疟疾诊断工具与检测方法的进展，重点比较了这些先进诊断技术在低收入/中低收入国家与发达经济体的应用差异。研究检索了 Google Scholar、PUBMED、多学科数字出版机构（MDPI）等科学数据库。结果表明：从快速诊断检测（RDTs）、分子检测技术到先进的无创检测方法和计算机化技术，疟疾检测领域已取得重要突破。值得注意的是，分子检测、RDTs 和计算机化检测在资源有限地区也已投入实际应用。 在总共八十个中低收入国家中，仅有二十一个国家（26%）显示出采用了现代疟疾诊断方法的证据。各国政府及相关机构亟需将工作重点转向疟疾研究，以加速全球消除疟疾的进程，特别是在通常资源有限的疟疾流行地区。

1. Introduction  1. 引言

Malaria elimination has been a focal topic of public health discussions for the past decade or more. Despite being a tropically endemic parasitic infection, the impact of malaria is far reaching and remains a global health concern. The 2023 World Health Organization (WHO) report states that malaria cases rose to an estimated 249 million in 2022, with an increase of 5 million more cases from the year 2021 [1]. Although relentless efforts are being made and strategies put in place, much more is required to free our globe of the parasitic infection, particularly in indigenous malaria-endemic countries such as a number of sub-Saharan African countries where most cases occur [2].
疟疾消除已成为过去十余年公共卫生讨论的核心议题。尽管属于热带地区流行的寄生虫感染，疟疾的影响却极为深远，至今仍是全球性健康威胁。世界卫生组织 2023 年度报告显示，2022 年全球疟疾病例数攀升至约 2.49 亿例，较 2021 年新增 500 万病例[1]。虽然各国持续实施防控策略并付出不懈努力，但要实现全球范围内（尤其是撒哈拉以南非洲等疟疾流行区本土国家——这些地区集中了绝大多数病例[2]）彻底消除这种寄生虫感染，仍需付出更多努力。

Central to eliminating malaria is early, accurate detection, quantification, and differentiation of the parasitic infection, especially among asymptomatic persons. Asymptomatic plasmodium-infected individuals represent a major threat to malaria elimination worldwide as they do not show signs of clinical disease yet serve as parasite reservoirs and significantly contribute to the spread of the infection [3]. Notably, the majority of these asymptomatic infections are missed by conventional diagnostic techniques. As a result, the need for reliable, sensitive, and specific diagnostic or detection methods arises, which would also be useful for monitoring any decline in malaria transmission [4].
消除疟疾的核心在于对这种寄生虫感染进行早期、准确的检测、定量和鉴别，尤其是在无症状人群中。无症状的疟原虫感染者虽未表现出临床症状，却作为寄生虫储存库存在，对疟疾在全球范围内的传播构成重大威胁[3]。值得注意的是，传统诊断技术往往会漏诊大多数这类无症状感染。因此，亟需建立可靠、灵敏且特异的诊断或检测方法，这些方法也将有助于监测疟疾传播的下降趋势[4]。

Technologies for malaria diagnosis have advanced in recent years; however, certain factors, such as the lack of laboratory infrastructure, operational costs, electricity requirements, and special operation expertise, have impeded the implementation of these advanced techniques in the vast majority of malaria endemic areas. This is especially the case when it comes to molecular testing, as these tests can be particularly expensive in addition to other challenges not only for malaria but for other infectious diseases [5]. The WHO describes microscopy (thin and thick film) as the primary method of detection [6]. Though microscopy is extensively used, it is unable to adequately detect low parasitemia, which is essential for effective treatment and subsequent elimination of the parasitic infection [7]. In addition, it is a laborious process requiring much expertise and experience for accurate diagnosis [4,8]. Other concerns have been the invasive approach of this technique, where blood samples are collected after a painful pierce of a needle, and yet an accurate diagnosis unassuredly relies solely on the discretion of the laboratory scientist. In several developing countries, there is inadequate expertise, equipment, and supplies required for accurate detection; as such, there are greater risks of contamination and false diagnosis [9]. Furthermore, it becomes more unreliable and difficult to distinguish low level infections as transmissions decline; hence, there is a need for alternative approaches to detection as elimination is being considered [4].
近年来，疟疾诊断技术虽取得进展，但实验室基础设施匮乏、运营成本高昂、电力供应需求以及专业操作技能等因素，阻碍了这些先进技术在绝大多数疟疾流行地区的推广应用。分子检测技术尤其如此，这类检测不仅费用特别昂贵，还面临疟疾及其他传染病共有的多重挑战[5]。世界卫生组织将显微镜检查（薄/厚血膜法）列为首要检测方法[6]。尽管显微镜检查应用广泛，却无法有效检出低原虫血症——这对实现有效治疗及后续消除寄生虫感染至关重要[7]。此外，该检测流程繁琐，需要大量专业知识和经验才能确保诊断准确性[4,8]。其他争议还包括该技术的侵入性操作（需通过疼痛的针刺采集血样），而准确诊断却仅依赖于实验室人员的主观判断。 在多个发展中国家，缺乏准确检测所需的专业知识、设备和物资；因此，存在更高的污染和误诊风险[9]。此外，随着传播率下降，区分低水平感染变得更加不可靠和困难；因此，在考虑消除疟疾时，需要采用替代检测方法[4]。

Can there be a faster, more specific, and more sensitive method of detecting malaria that can easily be implemented in resource-limited areas? The question remains among scientists globally. Can malaria be eliminated and many more lives saved by the emergence of technologies that offer early detection and differentiation of very minimal malarial infections? Does mankind stand a chance of advancement towards needle-free malaria detection, point-of-care devices, and personalized malaria medicine? For lower and lower-middle-income countries, which total 26 and 54, respectively (Table 1), according to the World Bank, will there be access to such effective diagnostic tools [10]? It is worthy of note that 11 of these countries, all in sub-Saharan Africa, bear 70% of the global malaria burden, according to the 2023 WHO report [1] (Table 1). The above-mentioned questions are but a few that remain on the minds of scientists and thus drive research.
是否存在一种更快、更特异且更灵敏的疟疾检测方法，能够轻松在资源有限地区实施？这仍是全球科学家共同关注的问题。随着能够早期检测和区分极微量疟疾感染技术的出现，人类是否有望消除疟疾并挽救更多生命？人类是否可能实现无针疟疾检测、即时诊断设备和个性化疟疾药物的突破？根据世界银行数据，对于分别有 26 个和 54 个的低收入及中低收入国家（表 1），这些地区能否获得此类高效诊断工具[10]？值得注意的是，根据 2023 年世卫组织报告[1]（表 1），其中 11 个全部位于撒哈拉以南非洲的国家承担着全球 70%的疟疾负担。上述问题仅是萦绕在科学家心头、推动相关研究的诸多疑问中的一小部分。

Table 1. Countries classified by World Bank as low or lower-middle-income economies in 2024 [1,10]. The table lists all resource-limited countries divided into low (top portion) and lower-middle (bottom portion) income countries, with special emphasis placed on the 11 countries (right portion) that together bear 70% of the global malaria burden.

	LOW AND LOWER-MIDDLE INCOME COUNTRIES
								70% GLOBAL MALARIA BURDEN
LOW-INCOME COUNTRIES	Afghanistan	Burundi	Central African Republic	Chad	Eritrea	Ethiopia	Gambia	Burkina Faso	Congo, Dem. Rep
	Guinea-Bissau	Korea, Dem. People’s Rep	Liberia	Madagascar	Malawi	Rwanda	Sierra Leone	Mali	Mozambique
	Somalia	South Sudan	Sudan	Syrian Arab Republic	Togo	Yemen, Rep		Niger	Uganda
LOWER-MIDDLE INCOME COUNTRIES	Angola	Algeria	Bangladesh	Benin	Bhutan	Bolivia	Cabo Verde	Cameroon	Ghana
	Cambodia	Comoros	Congo, Rep.	Côte d’Ivoire	Djibouti	Egypt, Arab Rep.	Eswatini	India	Nigeria
	Guinea	Haiti	Honduras	Jordan	Iran, Islamic Rep	Kenya	Kiribati	Tanzania
	Kyrgyz Republic	Lao PDR	Lebanon	Lesotho	Mauritania	Micronesia, Fed. Sts.	Mongolia
	Morocco	Myanmar	Nepal	Nicaragua	Pakistan	Papua New Guinea	Philippines
	Samoa	São Tomé and Principe	Senegal	Solomon Islands	Sri Lanka	Tajikistan	Timor-Leste
	Tunisia	Ukraine	Uzbekistan	Vanuatu	Vietnam	Zambia	Zimbabwe

Subsequently, there are several techniques that have been developed over the years to address some of the challenges with the gold standard technique. Rapid diagnostic tests (RDTs) are fast and reliable. Malaria RDTs do not require skilled personnel or constant electricity, but relative to malaria microscopy, they are expensive, have a short shelf life, and only give qualitative results [11]. Other diagnostic techniques, such as enzyme-linked immunosorbent assay (ELISA), lateral flow immunoassay (LFIA), microarrays, aptamer-based biosensors, genomic sequencing, loop-mediated isothermal amplification (LAMP), nested PCR, real-time PCR, and quantitative nucleic sequence-based amplification, are usually reserved for research and surveillance purposes. These techniques have higher sensitivity and specificity for malaria diagnosis relative to microscopy and RDTs. Notwithstanding, some of them are more laborious and expensive to deploy in resource-limited areas.
随后，多年来已开发出多种技术以应对金标准技术中的部分挑战。快速诊断检测（RDTs）具有快速可靠的特点。疟疾 RDTs 无需专业人员操作或持续电力供应，但与疟疾显微镜检查相比，其成本较高、保质期较短且仅能提供定性结果[11]。其他诊断技术如酶联免疫吸附试验（ELISA）、侧流免疫层析（LFIA）、微阵列、适配体生物传感器、基因组测序、环介导等温扩增（LAMP）、巢式 PCR、实时荧光定量 PCR 以及基于定量核酸序列的扩增技术，通常仅用于研究和监测目的。相较于显微镜检查和 RDTs，这些技术在疟疾诊断方面具有更高的敏感性和特异性。尽管如此，其中部分技术在资源有限地区实施时仍存在操作繁琐且成本高昂的问题。

This systematic review looks at traditional and modern techniques in light of their main advantages and disadvantages, as well as the countries where they have been used, with emphasis placed on lower and lower-middle-income countries. Also, emphasis would be placed on molecular-based techniques and how common they are in resource-limited settings, which usually happen to be endemic to malaria.
本系统综述从主要优缺点及适用国家维度，对比分析了传统与现代疟疾诊断技术，重点关注中低收入国家的应用情况。同时将着重探讨分子诊断技术的普及程度——这类技术往往在资源有限地区（通常也是疟疾流行区）的应用现状。

2. Materials and Methods  2. 材料与方法

In conducting this systematic review, an accurate and authentic outcome was ensured by adherence to the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines. The review was registered in the open science framework database (https://doi.org/10.17605/OSF.IO/DV6Z3 (accessed on 28 June 2024)). Relevant details needed were obtained from articles published in journals and from databases up to 2022 since the search was done in early 2023. Key search phrases used for the search included “malaria detection methods”, “emerging technologies in malaria”, “recent advances in malaria detection or diagnosis”, “emerging methods in malaria diagnosis”, “traditional methods of malaria detection”, “Point of care devices for malaria detection”, “Non-invasive or needle-free malaria detection”, and “personalized malaria medicine”. Numerous articles were obtained from databases, journals, and other publishing sites, including Google Scholar, PUBMED, and MDPI databases.
为确保本系统综述结果的准确性与可靠性，研究团队严格遵循《系统评价与荟萃分析优先报告条目》（PRISMA）指南开展工作。本综述已在开放科学框架数据库注册（注册号：https://doi.org/10.17605/OSF.IO/DV6Z3，访问日期 2024 年 6 月 28 日）。由于文献检索工作完成于 2023 年初，所有必要数据均来自 2022 年及之前发表的期刊论文与数据库资料。检索采用的核心关键词包括："疟疾检测方法"、"疟疾新兴技术"、"疟疾检测/诊断最新进展"、"疟疾诊断新兴方法"、"传统疟疾检测手段"、"疟疾即时检测设备"、"无创/无针疟疾检测"以及"个性化疟疾诊疗"。通过谷歌学术、PUBMED 和 MDPI 等数据库及期刊平台，我们获取了大量相关文献资料。

From the databases, a total of 327 were identified. After the searches, the publications were sorted out to remove duplicates, and 20 publications were removed. The records were further screened to remove all incomplete, unpublished articles, and ineligible publications. With articles published in recent years under consideration, all accessible publications were considerable options, leaving out articles from journals that needed to be purchased, were restricted, or there was not a PDF version of the complete paper readily available.
从各数据库中共识别出 327 篇文献。经过检索后，对出版物进行整理以去除重复项，移除了 20 篇文献。进一步筛选记录以排除所有不完整、未发表的文章以及不符合条件的出版物。考虑到近年发表的文章，所有可获取的出版物均被列为备选，排除了需要购买、受限制或无法直接获取完整论文 PDF 版本的期刊文章。

Upon abstract screening, articles were selected based on the following general criteria: a traditional or modern method of malaria detection or diagnosis was investigated. A total of 276 articles were obtained, uploaded into Mendeley Reference Manager and Endnote, and carefully reviewed for full text eligibility and results presentation.
在摘要筛选阶段，根据以下通用标准选择文章：研究采用传统或现代疟疾检测诊断方法。共获得 276 篇文章，将其上传至 Mendeley 文献管理器和 Endnote 软件，并仔细审查全文资格与结果呈现情况。

Of these articles, some investigated traditional methods, while most investigated various modern methods. Some articles were also found useful as they investigated or reviewed “emerging technologies in malaria detection” or “advances in malaria detection”, among others, and thus were used in the other parts of the review writing. After a comprehensive document screening, 167 were later removed as it was found that they did not suit the review criteria, leaving 109 to be reviewed. These articles were removed for reasons including the papers being published earlier than 2014 (for those to be analyzed), they did not specifically investigate diagnostic tools for malaria, the article obtained was not the published version, and the research scope and contents were not clear or did not focus on a possible malaria detection method. In several instances, multiple malaria detection methods were identified in a single publication; thus, the number of developed methods identified exceeded the number of publications used. Figure 1 below shows the sorting-out process.
在这些文献中，部分研究探讨了传统检测方法，而大多数则聚焦于各类现代检测技术。另有部分文献因涉及"疟疾检测新兴技术"或"疟疾检测进展"等主题研究或综述，被用于本综述的其他章节。经过全面筛选后，167 篇文献因不符合综述标准被剔除（剔除原因包括：发表时间早于 2014 年[针对待分析文献]、未专门研究疟疾诊断工具、获取版本非最终发表稿、研究范围与内容不明确或未聚焦可行的疟疾检测方法），最终保留 109 篇文献进行评述。值得注意的是，部分单篇出版物中识别出多种疟疾检测方法，因此所鉴定的检测方法数量超过了实际采用的出版物数量。图 1 展示了文献筛选流程。

Figure 1. A PRISMA flow diagram showing the method of article selection.
图 1. 展示文献筛选方法的 PRISMA 流程图。

3. Results  3. 结果

3.1. Traditional Methods Used for Malaria Detection
3.1 疟疾检测的传统方法

Table 2 below shows a summary of traditionally used methods of malaria detection, elaborating on their approach as well as the pros and cons of using these methods for diagnosis. Though there has been the development of new and innovative methods of detection over the years, microscopy, using thick and thin blood films coupled with Giemsa staining, remains the gold standard for the diagnosis of malaria parasitic infections [8,12].
下表 2 总结了传统疟疾检测方法，详细说明了这些方法的操作流程及其诊断应用的优缺点。尽管近年来已开发出诸多创新检测技术，但采用厚薄血片结合吉姆萨染色的显微镜检查法，仍是诊断疟原虫感染的金标准[8,12]。

Table 2. Traditional methods used for malaria detection.
表 2. 疟疾检测使用的传统方法

Traditional Methods  传统方法	Specimen Used  使用样本	Summary of Procedure  操作步骤概述	Invasive/Non-Invasive  侵入性/非侵入性	Advantages  优势	Disadvantages  劣势	Refer-ences  参考文献
Thin film microscopy  薄片显微镜检查	Blood  血液	Thin blood smears are prepared and stained using Giemsa stain. Thin smears are examined with a 100× oil immersion objective. 薄血涂片采用吉姆萨染色法制备。薄涂片需使用 100 倍油镜进行镜检。	Invasive  侵入性	Reliable in the identification of four human plasmodium species and their various stages 可准确鉴别四种人类疟原虫及其各发育阶段	Limited by quality of blood smears as well as availability of skilled microscopists. 受限于血涂片质量及熟练显微镜技师的稀缺性。 Lack of sensitivity where non-falciparum or mixed infections exist. 对非恶性疟原虫或混合感染检测灵敏度不足。	[8,13,14,15,16,17,18]
Thick film microscopy  厚血膜镜检法	Blood  血液	Thick blood smears are prepared and stained using Giemsa stain. Thin smears are examined with a 100× oil immersion objective. 厚血涂片采用吉姆萨染色法制备。薄血涂片需使用 100 倍油镜进行观察。	Invasive  侵入性检测	Reliable in the detection of four human plasmodium species 可可靠检出四种人类疟原虫	Limited by quality of blood smears as well as availability of skilled microscopists. 受限于血涂片质量及熟练显微镜技师的稀缺性	[8,13,14,15,16,17,18]
Morphology-based diagnosis 基于形态学的诊断	Blood  血液	Optical images from Giemsa-stained infected blood are measured using Olysia and Scanning Probe Image Processor software based on morphology of red blood cells. 通过 Olysia 和扫描探针图像处理器软件，基于红细胞形态对吉姆萨染色感染血液的光学图像进行测量。	Invasive  侵入性	Faster prediction of malaria cases 更快预测疟疾病例	Expertise needed  需要专业知识	[19]  《资源有限地区疟疾诊断方法进展：系统性综述》
Centrifuged buffy coat smear examination (CBCS) 离心后白细胞层涂片检查（CBCS）	Blood  血液	Centrifugation of buffy coat is done prior to Giemsa staining and microscopic examination 在吉姆萨染色和显微镜检查前需进行血沉棕黄层离心处理	Invasive  侵入性检测	Specificity is similar to conventional method but sensitivity a bit better than conventional method 特异性与传统方法相当，但灵敏度略优于传统方法	Limited by availability of skilled microscopists 受限于熟练显微镜操作人员的稀缺性	[20]

3.2. Modern Methods Used for Malaria Detection
3.2. 资源有限地区疟疾诊断方法进展：系统综述

The quest to effectively treat malaria while gravitating towards its elimination has driven the development of various tools and assays for the diagnosis of malaria (4). Table 3 and Table 4 contain recently developed methods used in the diagnosis of malaria and where they have been used. These diagnostic approaches vary greatly, ranging from biosensors and molecular assays down to computerized algorithms and automated analyzers, which have been developed or used over recent years, no earlier than 2014. The advantages and limitations of each diagnostic method are considered, as well as the summarized procedure by which it is conducted.
在有效治疗疟疾并逐步实现消除疟疾的目标驱动下，各种疟疾诊断工具和检测方法得以发展（4）。表 3 和表 4 列出了近年来（不早于 2014 年）开发或应用的疟疾诊断新方法及其使用地区。这些诊断方法差异显著，涵盖从生物传感器、分子检测到计算机算法和自动化分析仪等多种技术。每种诊断方法的优势与局限性均被纳入考量，并概述了其实施流程。

Table 3. Modern (PCR/LAMP-based) methods used for malaria detection and evidence of use in developed countries.

Modern Methods	Specimen Used	Description	Invasive/Non-Invasive	Point of Care/Molecular/Other	Advantages	Disadvantages	Developed Countries	References
Direct conventional PCR	Blood	With plasmodium cytochrome oxidase III gene (COX-III) as target, direct conventional PCR is conducted on bloodspot samples. Results are visualized on a gel.	Invasive	Molecular	High Sensitivity; faster than nested; does not require DNA isolation	Requires much expertise and expensive	USA	[21]
Nested Polymerase Chain Reaction (PCR)	Blood	Using different primer pairs to run 2 sequential amplification reactions. Plasmodium genomic DNA extracted from dried blood spots	Invasive	Molecular	High sensitivity and specificity	Time consuming, expensive, requires much expertise	Thailand, USA, Brazil, United Kingdom, Austria	[13,16,18,21,22,23,24,25]
Droplet Digital PCR (ddPCR)	Blood, Serum	DNA extracted from blood and serum samples are analyzed using the ddPCR method, which is based on water–oil emulsion droplet technology	Invasive	Molecular	High sensitivity using blood samples	Low sensitivity using serum samples; expensive	Italy, Thailand	[26,27]
Photo- Induced Electron transfer PCR (PET-PCR)	Blood	Total DNA is extracted from dried blood spots and PCR performed using photo-induced electron transfer fluorogenic primers	Invasive	Molecular	High sen-sitivity for parasite identification and characterization.	Requires much expertise and is expensive	USA	[15]
Fluoresen-ce reson-ance energy transfer (FRET) real time PCR	Blood	Real-time PCR utilizing FRET whereby fluorophores are brought in close proximity after hybridization is performed on DNA extracted from lyophilized blood samples targeting the 18S rRNA gene	Invasive	Molecular	High sensit-ivity, and allows for simultaneous quantitative and species-specific detection	This specific protocol could not differentiate between P. vivax and P. knowlesi; expensive	United Kingdom, Austria	[22]
SYBR Green Real-Time PCR-RFLP Assay	Blood	Real-time PCR using sybr green dye that binds to all double-stranded DNA followed by restriction fragment polymorphism to differentiate species	Invasive	Molecular	High sensitivity	Meltcurve required in PCR since Sybr green alone can be non-specific; expensive	Sweden	[28]
Hair qPCR	Head hairs	Hairs without roots are taken from patients and qPCR assay conducted	Non-invasive	molecular	Requires no special trans-port/storage conditions for samples	Sensitivity lower than when blood samples are used	Spain	[29]
Insulated Isothermal PCR (iiPCR)	Blood	PCR is performed in a portable device using an assay based on the Rayleigh–Bénard convection method	Invasive	Molecular/point of care	Portable, easy and fast operation; direct interpretation	Not as sensitive as qPCR	Malaysia	[30]
Lab Chip Real Time PCR (LRP)	Blood	DNA is extracted from collected blood samples and a portable LRP device is used to detect malarial parasites based on lab-on-chip technology	Invasive	Molecular/point of care	High sensitivity and specificity. Fast and cost effective	Risk of false negatives higher than traditional real-time PCR	Korea	[31]
Pv-mt Cox PCR	Blood	DNA is extracted from collected blood samples and qPCR with mitochondrial gene target is carried out	Invasive	Molecular	More sensitive in the detection of P. vivax	Expensive	Brazil	[32]
PvLAP5 and Pvs25qRT-PCR assays	Blood	Extracted RNA is subjected to quantitative reverse transcription PCR	Invasive	Molecular	Suitable assay for the determination of human transmission reservoir	Expensive	Panama	[33]
Other Quantita-tive PCR (qPCR)	Blood	Real-time PCR performed using primers targeting different regions and SYBR green or probe-based technology on DNA extracted from whole blood	Invasive	Molecular	High sensitivity and rapid	Extreme caution needed to prevent contamination; expensive	France, Canada, USA Columbia Germany, Brazil, China, Malaysia	[34,35,36,37,38,39,40,41,42,43,44]
Dry LAMP system (CZC-LAMP)	Blood	Blood samples are analyzed directly without extraction using the assay made up of dried reagents	Invasive	Point of care/molecular	High sensitivity and specificity; no need for prior extraction	Not widely available		[45]
Particle Diffusometry (PD)-LAMP	Blood	PD, which senses images, is combined with LAMP on a smartphone-enabled device to detect low levels of parasitemia	Invasive	Point of care/molecular	Sensitivitities higher than RDTs and similar to qPCR	Sensitivity slightly lower than nested PCR	USA	[46]
LAMP and MinION™ nanopore sequencer	Blood	Primers targeting the 18S–rRNA gene of all five human Plasmodium species are generated and samples subjected to LAMP. Min-ION™ nanopore sequencer is used on amplicons to identify Plasmodium spp.	Invasive	Molecular	Highly sensitive, and simple	Expensive	Japan	[47]
Other Loop-mediated isothermal amplification (LAMP),	Blood	Extracted DNA is subjected to loop-mediated isothermal amplification with a variety of detection methods	Invasive	Point of care/molecular	Simple, low cost; can be used in resource-limited settings and point-of-care settings	Some cannot quantify par-asite density; some are insensitive towards low parasitemia and mixed infections	France, Korea, Thailand Italy, Brazil Spain, Mala-ysia, Japan, Peru, USA	[26,34,48,49,50,51,52,53,54,55,56,57,58,59,60,61,62,63]

Table 4. Modern (non-PCR/non-LAMP-based) methods used for malaria detection and evidence of use in developed countries.

Modern Methods	Specimen Used	Description	Invasive/Non-Invasive	Point of Care/Molecular/Other	Advantages	Disadvantages	Developed Countries	References
Malaria SD Bioline RDT kit	Urine, Saliva, Blood	Using immuno-chromatography to detect PfhRP2 and PLDH following manufacturer’s instructions	Non-invasive/Invasive	Point of care	Effective for non-invasive detection of malaria; low cost	Low sensitivity		[64]
Other (RDTs)	Blood	Immunochromatography/ according to manufacturer’s instructions	Invasive	Point of care	Suitable for point of care in hard-to-access areas; low cost	Low sensit-ivity for some kits; poor identification of non-falciparum infections for some brands	Indonesia Australia, USA	[14,15,17,18,65,66,67,68,69,70,71]
Computeri-zed/digital deep mach-ine learnin-g approach	Blood	Machine learning models are used to detect malaria parasites in blood smears. Some can be integrated into smartphone detection apps	Invasive	Other	Accurate/ reliable	For some, results are affected by quality of smears	USA, Taiwan, China, Turkey	[72,73,74,75,76,77,78,79,80,81]
Spectros-copy	Blood	Blood samples are analyzed using spectroscopy	Invasive	Other	Highly effective for identifying infected cell	Only qualitative results obtained	Thailand, China, Australia	[82,83,84]
Portable Optical Diagnostic System (PODS)	Blood	Works by differential optical spectroscopy. The change in optical power before and after a magnet is applied, is monitored in order to determine β-hematin concentration in whole blood	Invasive	Point of care	Portable; low cost; useful for low resource settings; high sensitivity	Not widely available	USA	[85]
Ultra bright SERS nanorattles	Blood	DNA detection method that uses sandwich hybridization of magnetic bead, target sequence, and ultrabright SERS nanorattle are employed	Invasive	Molecular/point of care	Sensitive; can be automated and added to portable devi-ces for POC diagnosis; can identify SNPs hence, discri-minate betw-een wild-type and mutant parasites	Not widely available	USA	[86]
Automated Microscopy/Digital Analysis	Blood	Comprises a fluorescent dye staining or Giemsa staining and an automated microscopy platform and digital analysis	Invasive	Other	Rapid diagn-osis and par-asite density monitoring. High sens- itivity, linear-ity, and precision	Not widely available	Korea, Finland, Sweden	[87,88,89]
Flow cytometry	Blood	Parasites are detected and quantified in blood by use of analyzers utilizing flow cytometry technology	Invasive	Molecular	Rapid and high sensiti-vity; useful for mass screening	May not be able to distinguish plasmodium species	Netherlands, France, USA, South Africa, Japan	[90,91,92,93,94]
Thin-Film Optical Filters	Blood	A thin film optical device is used based on optical reflectance spectrophotometry, for the parasite detection through haemozoin quantification	Invasive	Point of care	High sensitivity	High transmittance regions outside target wavelength	Portugal	[95]
Rotating cr- ystal magn-eto optical detection (RMOD) method	Blood	RMOD works by detection of the periodic modulation of light transmission. This is induced by hemozoin crystals which co-rotates with a rotating magnetic field	Invasive	Other	Higher sensitivity and accuracy than light microscopy	Sensitivity is poorer than PCR methods	Thailand, Hungary	[96,97,98]
Hemozin-Based Malaria diagnostic device (GazelleTM)	Blood	Using magneto-optical technology, the device detects hemozoin produced by Plasmodium	Invasive	Other	Sensitivities comparable to light micr-oscopy; faster than micros-copy; portab-le; can run on battery power	Unable to distinguish between species		[16]
Hemozoin-generated vapor nanobubbles	Blood vessel (transdermal)	Hemozoin generates a transient vapor nanobubble around hemozoin in response to a short and safe laser pulse. The acoustic signals of these nanobubbles that are malaria specific enable detection	Non-invasive	Point of care	Non-invasive; rapid	Not widely available	USA	[99]
Electroche-mical immunosensor	Blood	Egg yolk IgY antibodies against Plasmodium vivax lactate dehydrogenase antigen are immobilized on a gold electrode surface followed by differential pulse voltammetry and contact angle measurements are made.	Invasive	Point of care	High Sensitivity for malaria caused by P. vivax	Only malaria caused by P. vivax can be detected	Brazil	[100]
Simplified ELISA)/PfHRP 2 ELISA	Blood	Modified ElISA was performed on blood samples.	Invasive	Point of care	High sensitivity, portable and low cost	Not widely available	Spain UK Denmark	[101,102]
Multiple-xed ELISA based assay	Blood	Multiplexed ELISA-based (either planar-based array or magnetic bead-based platforms) technologies are used for malaria detection	Invasive	Molecular	Can detect malaria spe-cies mutants; have high throughput potential	Not widely available	USA	[103]
Dye-Cou-pledApt-amer-Capt-ured Enzy-me-Cataly-zed assay	Blood	Aptamer- and enzyme-based method is used to detect malaria infection in blood. Method can be used on instrument or instrument free platform	Invasive	Molecular/point of care	Low cost; useful for resource-limited and point-of-care settings.	Not widely available		[104]
Recombinase-Aided Amplificat-ion with Lateral Flow Dip-stick Assay (RAA-LFD)	Blood	A combination of recombinase-aided amplification lasting for 15 min at 37 degrees and lateral flow dipstick is used to detect plasmodium species in blood	Invasive	Molecular/point of care	Highly sensitive, specific, low cost, convenient for on-site screening and low resource settings.	Not widely available	China	[105]
Portable image-based Cytometer	Blood	P. falciparum-infected blood cells are identified and counted from Giemsa-stained smears using the image based portable cytometer.	Invasive	Other	Simple to operate; low cost	Not widely available	Singapore	[106]
Two-stage sample-to-answer sy-stem based on nucleic acid ampl-ification approach	Blood	It combines the dimethyl adipimidate (DMA)/thin film sample processing (DTS) technique and the Mach–Zehnder interferometer isothermal solid-phase DNA amplification (MZI-IDA) technique to detect infection in blood	Invasive	Molecular	High sensitivity, rapid	Not widely available	Singapore, Korea	[107]
Fluorescen-ce In Situ Hybridization (FISH) Assays	Blood	Detects and localizes specific malaria nucleic acid sequences by hybridizing with complementary sequences that are labeled with fluorescent probes	Invasive	Molecular	High sensitivity	Skilled expertise required.	USA	[108,109]
NMR-based hemozoin detection	Blood	Detection is based on the ability to recognize the paramagnetic susceptibility of malaria hemozoin crystals	Invasive	Molecular/point of care	High sensitivity and rapid	Not widely available	Australia, Singapore, USA	[110,111,112]
Multi-omics	Varies	Integrating data from different omic methods	Invasive/non-invasive	Other	Comprehen-sive underst-anding of the infection	Requires skilled experitise	Austria USA Columbia	[113,114,115,116]

Table 5 analyzes evidence of the use of some recently developed detection tools in lower and lower-middle-income countries where there are often resource limitations. The test types that featured most frequently in publications were PCR techniques (eleven), followed by RDT tests (nine), then LAMP techniques and computerized/digital deep machine learning approaches (six each). In all, twenty-one countries had publications featuring modern malaria diagnostic methods.
表 5 分析了在资源往往有限的中低收入国家使用一些最新开发的检测工具的证据。出版物中最常出现的检测类型是 PCR 技术（11 项），其次是 RDT 检测（9 项），然后是 LAMP 技术和计算机化/数字深度学习技术（各 6 项）。总共有 21 个国家发表了采用现代疟疾诊断方法的出版物。

Table 5. Evidence of use of modern methods of malaria detection in low and lower-middle-income countries.

Modern Method	Resource-Limited Countries	References
Malaria rapid test kit (SD Bioline RDT kit) using urine and saliva samples	Ghana	[64]
Other rapid diagnostic tests	Nigeria, Senegal, Kenya, Benin, Pakistan, Burkina Faso	[14,15,17,18,65,66,68,69]
Nested polymerase chain reaction (PCR)	Pakistan, Nigeria, Myanmar, Honduras, India	[13,16,18,23,25]
Hair qPCR	Rwanda	[29]
Other quantitative polymerase chain reaction (qPCR)	Bangladesh, Eritrea, Tanzania D.R. Congo, Sierra Leone, Cambodia	[35,36,37,38,40]
Dry LAMP system (CZC-LAMP	Zambia	[45]
Other loop-mediated isothermal amplification (LAMP),	India, Tanzania, Senegal, Ghana	[48,56,57,58,59]
Computerized/digital deep machine learning approach	Nigeria, Uganda, Bangladesh, Ethiopia, Zambia,	[59,75,77,78,79,80]
The rotating-crystal magneto-optical detection (RMOD) method	Papua New Guinea	[96]
Hemozin-based malaria diagnostic device (GazelleTM)	Honduras	[16]
Flow cytometry	Burkina Faso, India	[90,93]
Dye-coupled aptamer-captured enzyme-catalyzed assay	India	[104]
Multi-omics	India	[114]

In Figure 2, the various methods of malaria detection reported from the identified studies have been represented graphically, indicating which diagnostic trends are being largely investigated, used more, or have gained much research interest. The chart represents malaria diagnostic developments investigated from 2014 until 2022. PCR-based methods and LAMP-based methods were the most prevalent methods.
图 2 以图形方式呈现了所纳入研究中报告的各种疟疾检测方法，展示了哪些诊断趋势受到广泛研究、更频繁使用或获得较多研究关注。该图表反映了 2014 年至 2022 年间研究的疟疾诊断技术发展情况。基于 PCR 的方法和基于 LAMP 的方法是最主流的检测技术。

Figure 2. Frequencies of malaria detection/diagnosis methods in reviewed publications.
图 2. 已综述文献中疟疾检测/诊断方法的使用频率。

4. Discussion  4. 讨论

Critical to achieving effective control, treatment, and subsequent elimination of malaria is the timely detection of the parasitic infection. In the face of this threatening infection, continuous progress and innovative research are required, which leads to the development of new tools that will be useful in the fight against malaria [117]. This article reviewed the recent developments in malaria diagnostic methods and their potential for point-of-care and personalized malaria care, with special emphasis on the use of these methods in economically challenged countries.
实现疟疾有效控制、治疗及最终消除的关键在于及时检测寄生虫感染。面对这一威胁性感染，需要持续进展与创新研究，从而开发出有助于抗击疟疾的新工具[117]。本文综述了疟疾诊断方法的最新进展及其在即时检测和个性化疟疾护理中的应用潜力，特别关注了这些方法在经济欠发达国家中的使用情况。

The findings from this review suggest great advancement recently in malaria diagnostics. Research efforts by many scientists around the globe have progressed from developing improved malaria microscopy techniques into enhanced and more accurate molecular, immunological, computerized, digital methods of detection, automated analyzers, and point-of-care devices. Studies suggest that the influence of the old age infection on global health outcomes has urged on the design of more efficient diagnostics, with efforts directed at the development of point-of-care devices useful for resource-limited areas [7]. For an active drive towards the elimination of malaria, an early detection approach capable of revealing low levels of the parasitic infection is imperative [3].
本综述的研究结果表明，近期疟疾诊断技术取得了重大进展。全球众多科学家的研究已从改进疟疾显微镜技术，发展到开发更精准的分子、免疫学、计算机化、数字化检测方法，以及自动化分析仪和即时检测设备。研究表明，这种古老传染病对全球健康的影响促使人们设计出更高效的诊断方法，重点开发适用于资源有限地区的即时检测设备[7]。要实现消除疟疾的积极目标，必须采用能够检测低水平寄生虫感染的早期诊断方法[3]。

As observed in Table 2, Table 3, Table 4 and Table 5, the outcome of this review indicates that recent malaria detection methods actively being used or investigated include traditional methods, molecular techniques with polymerase chain reaction (PCR), loop-mediated isothermal amplification (LAMP)-based assays, and machine learning/computerized techniques (that exploit the physical and/or biological properties of Plasmodium-infected erythrocytes to enhance malaria diagnosis), among others. Figure 2 shows the frequencies of detection methods as identified from various articles published over the last decade. Several other technologies and chemical assays are also being designed to tackle the malaria burden. RDTs were among the commonly used or researched modern methods in resource-limited settings, as seen in Table 5 and Figure 2. This is not surprising since they are relatively low cost and easy to use.
如表 2、表 3、表 4 和表 5 所示，本综述结果表明当前正在使用或研究的疟疾检测方法包括传统方法、基于聚合酶链反应（PCR）的分子技术、环介导等温扩增（LAMP）检测法，以及机器学习/计算机化技术（利用疟原虫感染红细胞的物理和/或生物学特性来增强疟疾诊断）等。图 2 展示了近十年发表文献中各类检测方法的使用频率。目前还有多项其他技术和化学检测法正在研发中以应对疟疾负担。如表 5 和图 2 所示，在资源有限地区，快速诊断检测（RDTs）属于常用或重点研究的现代方法之一，这并不令人意外，因其成本相对较低且易于操作。

Studies confirm PCR-based techniques as having widespread use globally as they are highly sensitive and capable of detecting very low parasitemia levels [3,22,35]. Polymerase chain reaction basically makes use of DNA extracted from whole blood or other samples. The process continues with denaturation, amplification, and elongation steps, after which the sensitivity and specificity of the assay can be assessed [35]. It is proposed that the PCR method, under equal reaction parameters, can diagnose all five species of the plasmodium parasitic infection [35]. Our findings reveal that a wide range of PCR assays have been developed or used over the past decade, which are less laborious and provide much faster and more accurate results [22]. Furthermore, PCR-based assays are widely preferred due to several reasons, including simultaneous species-specific detection and quantification, higher sensitivity, higher specificity, less time consuming, easy to use, and capable of diagnosing subclinical infections [13,18,22,23,41,42].
研究证实基于 PCR 的技术因其高灵敏度且能检测极低寄生虫血症水平而在全球广泛应用[3,22,35]。聚合酶链式反应主要利用从全血或其他样本中提取的 DNA 进行操作。该过程包括变性、扩增和延伸步骤，之后可评估检测的灵敏度和特异性[35]。研究表明在相同反应参数下，PCR 方法可诊断所有五种疟原虫寄生感染[35]。我们的研究结果显示过去十年已开发或使用了多种 PCR 检测方法，这些方法操作简便且能提供更快速准确的结果[22]。此外，基于 PCR 的检测方法因多项优势被广泛采用，包括可同步进行虫种特异性检测与定量分析、更高灵敏度、更强特异性、耗时更少、操作简便以及能诊断亚临床感染等[13,18,22,23,41,42]。

Though PCR is an effective approach to malaria detection, it is limited by the requirement of costly laboratory facilities and expertise and thus less beneficial to resource-limited areas and at the point of care [3]. Despite that, quite a number of studies in resource-limited settings, including some African countries, utilized PCR-based techniques, as shown in Table 5 and Figure 2 [13,16,18,23,25,29,35,36,37,38,40]. Other advanced PCR techniques, such as lab chip real-time PCR (LRP) and hair qPCR, were found to be suitable alternatives for point-of-care or resource-limited settings, though no evidence was found of the former currently being used or researched in lower or lower-middle-income countries [29,31]. Gómez-Luque et al. proposed that due to limitations observed, more research is required to affirm the use of the hair qPCR as an efficient technique for malaria detection [29]. The one advantage the hair qPCR has over other PCR types is the use of non-invasive samples. LRP, however, being highly sensitive, specific, and less expensive will be beneficial for diagnosis and control in malaria-endemic countries [31].
尽管 PCR 是一种有效的疟疾检测方法，但由于需要昂贵的实验室设备和专业技术人员，其在资源有限地区和即时检测场景中的应用价值受限[3]。尽管如此，如表 5 和图 2 所示，包括部分非洲国家在内的资源有限地区仍有大量研究采用基于 PCR 的技术[13,16,18,23,25,29,35,36,37,38,40]。其他先进 PCR 技术（如实验室芯片实时 PCR（LRP）和毛发 qPCR）被发现适用于即时检测或资源有限环境，但目前尚未发现前者在低收入或中低收入国家应用或研究的证据[29,31]。Gómez-Luque 等学者指出，鉴于观察到的局限性，需要更多研究来确认毛发 qPCR 作为疟疾检测高效技术的可行性[29]。毛发 qPCR 相较于其他 PCR 类型的优势在于使用非侵入性样本。而 LRP 凭借高灵敏度、高特异性和较低成本，将为疟疾流行国家的诊断与防控提供重要帮助[31]。

LAMP-based assays have also dominated research on malaria diagnostics. As seen in Table 3, studies have shown the development or use of various LAMP assays, which are effective malaria diagnostics [118]. Rei Yan et al. reviewed LAMP assays and found them easy to use in regions where there is limited access to clinical expertise and molecular biology equipment. Modified LAMP based assays such as multiplex LAMP with dipstick DNA chromatography, high throughput LAMP, 18S rRNA LAMP, mediated LAMP combined with lateral flow detection (LFD), etc., are highly sensitive, easy to use, consistent, convenient, cost effective, and useful in point-of-care situations [55,56,58,59], thus enabling an approach towards personalized healthcare. Table 5 provides evidence of the development and use of LAMP techniques in lower and lower-middle-income countries, including countries in sub-Saharan Africa where malaria is endemic [45,48,56,57,58,59].
基于 LAMP 技术的检测方法在疟疾诊断研究中同样占据主导地位。如表 3 所示，多项研究证实了各类 LAMP 检测技术的开发与应用，这些技术都是有效的疟疾诊断手段[118]。Rei Yan 等学者对 LAMP 检测技术进行综述后发现，该技术在临床专业知识和分子生物学设备获取受限的地区易于实施。改良型 LAMP 检测技术——包括结合试纸条 DNA 色谱的多重 LAMP、高通量 LAMP、18S rRNA LAMP、联合侧流层析检测（LFD）的介导 LAMP 等——具有高灵敏度、操作简便、结果稳定、使用便捷、成本效益显著等优势，适用于即时检测场景[55,56,58,59]，为实现个性化医疗提供了技术路径。表 5 列举了 LAMP 技术在低收入和中低收入国家（包括疟疾流行的撒哈拉以南非洲地区）开发与应用的相关证据[45,48,56,57,58,59]。

Other molecular methods worthy of note as they double as point-of-care or easy-to-use methods include nuclear magnetic resonance (NMR)-based hemozoin detection, ultra-bright SERS nanorattles, recombinase-aided amplification with lateral flow dipstick assay, and dye-coupled aptamer-captured enzyme-catalyzed assay [86,104,105,110,111,112]. Though the latter two could be used in resource-limited settings due to their low cost, the study found only a dye-coupled aptamer-captured enzyme-catalyzed assay used in India [104]. Veiga and Peng identified nuclear magnetic resonance (NMR)-based hemozoin detection as having the potential of enabling personalized malaria medicine (that is, malaria treatment tailored to individual characteristics) with needleless diagnosis foresighted [119]. This technology may offer the detection of phenotypic variants, which are observable variations in characteristics among parasites of the same species as a result of genetic diversity, host–parasite interactions, or environmental factors, among others [120,121]. For example, there are drug-resistant variants, those with surface antigen variations, and variants with different clinical presentations, among others [121,122,123]. The ability to detect such variants would increase diagnostic accuracy and be considerably useful against parasite drug resistance. Acquiring these time- and patient-specific phenotypic identifiers is a basic step to personalized malaria medicine as variants continually rise [119]. The one advantage that phenotypic variant determination using NMR technology may have over nucleic acid amplification-based methods for genomic profiling is the extremely fast turnaround time for some of the devices [110]. Unfortunately, there was no evidence of such methods being used in lower and lower-middle-income countries as per the studied published data in the research articles reviewed.
其他值得注意的分子检测方法因其兼具即时检测或操作简便特性，包括基于核磁共振(NMR)的血红素检测、超亮表面增强拉曼散射纳米摇铃技术、重组酶辅助扩增结合侧流层析试纸条检测法以及染料偶联适体捕获酶催化分析法[86,104,105,110,111,112]。尽管后两种方法因成本低廉可用于资源有限地区，但研究发现仅染料偶联适体捕获酶催化分析法在印度得到应用[104]。Veiga 和 Peng 指出基于核磁共振的血红素检测技术有望实现无针诊断，为个性化疟疾治疗（即根据个体特征定制治疗方案）提供可能[119]。该技术可检测表型变异——由遗传多样性、宿主-寄生虫相互作用或环境因素等导致的同种寄生虫间可观察特征差异[120,121]。 例如，存在耐药变异株、表面抗原变异株以及具有不同临床表现的变异株等[121, 122, 123]。检测此类变异株的能力将提高诊断准确性，并对抗寄生虫耐药性具有重要价值。随着变异株持续增加，获取这些具有时间特异性和患者特异性的表型标识符是实现个性化疟疾治疗的基础步骤[119]。与基于核酸扩增的基因组分析方法相比，利用核磁共振技术进行表型变异检测的一个优势在于某些设备的检测速度极快[110]。遗憾的是，根据所综述研究文章中的已发表数据，目前没有证据表明此类方法在低收入和中低收入国家得到应用。

Furthermore, a number of other technologies have emerged capable of point-of-care diagnosis. Unlike the traditional microscopy and commonly used RDTs, some of these methods were found to be highly sensitive, non-invasive as far as sample collection was concerned, and cost effective, even though there was no evidence that cost-effective ones were necessarily being used in economically challenged settings [85,95,99,100,101,102]. In addition to these, Aggarwal et al. classify omics-based diagnostics as another important category to malaria diagnosis and elimination [124]. Multi-omics combines genomics, proteomics, metabolomics, phenomics, and transcriptomics in the investigation of biomarkers optimal for disease diagnosis and treatment. Though each omics has individual limitations, collectively, multi-omics can lead to a more comprehensive understanding of malaria infections, which can lead to more effective treatments [124]. In this review, the only struggling economy we found using multi-omics was India. No African nation was indicated.
此外，多种可实现即时诊断的新技术相继涌现。与传统显微镜检测和常用快速诊断试纸不同，部分新技术被证实具有高灵敏度、样本采集无创性以及成本效益优势，尽管尚无证据表明这些高性价比方法已在经济欠发达地区得到应用[85,95,99,100,101,102]。除上述技术外，Aggarwal 等学者将组学诊断技术归类为疟疾诊断与消除领域的另一重要范畴[124]。多组学技术通过整合基因组学、蛋白质组学、代谢组学、表型组学和转录组学，筛选最适合疾病诊断与治疗的生物标志物。尽管各类组学技术均存在局限性，但多组学联用能更全面地解析疟疾感染机制，从而催生更有效的治疗方案[124]。本综述发现，印度是唯一应用多组学技术的发展中经济体，未见非洲国家相关报道。

5. Conclusions  5. 结论

Given the literature reviewed, there is adequate evidence to suggest that malaria detection or diagnosis will progress significantly in the next decade and beyond towards needleless detection. This advancement will however require increased, detailed, and specified research into the various molecular identifiers and phenotypic variant characteristics of malaria infection while enhancing the accuracy, precision, and specificity of the modernized point-of-care diagnostic tools. With this in view, precedence is duly set for the use of personalized medicine in the treatment of malaria infections. Notwithstanding, the traditional thin and thick film microscopy and RDTs will continue to play an important role in the accurate detection of malaria infections, especially in resource-limited areas where there is less access to modernized diagnostic tools and little research into advanced malaria detection methods. It is, however, encouraging to see that PCR-based and LAMP-based tests were seen being utilized in these areas, including African countries. However, other modern molecular/point-of-care tests were not being utilized in sub-Saharan Africa. Findings of this study show that approximately a quarter (26%) of a total of eighty countries in low and lower-middle-income settings employ state-of-the-art methods for malaria diagnostics. This underscores the need for governments, non-governmental organizations, and funding bodies to intensify efforts towards malaria diagnostics and research in the fight against malaria.
根据文献综述，现有充分证据表明疟疾检测诊断技术将在未来十年及更长时间内朝着无创检测方向取得重大进展。然而这一进步需要对疟疾感染的各类分子标识物和表型变异特征开展更深入、细致且针对性的研究，同时提升现代化即时诊断工具的准确性、精密度和特异性。由此可见，个性化医疗在疟疾治疗中的应用已奠定基础。尽管如此，传统薄/厚血片镜检和快速诊断试纸条仍将在疟疾精准检测中发挥重要作用，特别是在医疗资源有限、现代化诊断工具匮乏且缺乏先进疟疾检测方法研究的地区。值得欣慰的是，包括非洲国家在内的这些地区已开始应用基于 PCR 和 LAMP 技术的检测方法，但撒哈拉以南非洲地区尚未推广其他现代化分子/即时检测技术。 本研究发现，在八十个低收入和中低收入国家中，约四分之一（26%）采用了最先进的疟疾诊断方法。这凸显出各国政府、非政府组织和资助机构亟需加强疟疾诊断和研究工作，以推进疟疾防治进程。