Amambo Glory Ngongeh, Innocentia Ngong, Abong Raphael Awah, Fombad Fanny Fri, Njouendou Abdel Jelil, Nietcho Franck, Ekanya Relindis, Kien Chi Anizette, Ebai Rene, Lenz Benjamin, Ritter Manuel, Esum Mathias Eyong, Deribe Kebede, Cho Jerome Fru, Beng Amuam Andrew, Enyong Peter Ivo, Li Zhiru, Hübner Marc P, Pfarr Kenneth, Hoerauf Achim, Carlow Clotilde, Wanji Samuel
Parasites and Vector Research Unit (PAVRU), Department of Microbiology and Parasitology, University of Buea, Buea, Cameroon.
Research Foundation in Tropical Diseases and Environment (REFOTDE), Buea, Cameroon.
Front Trop Dis. 2023 Jan 9;3:1016176. doi: 10.3389/fitd.2022.1016176.
Conventional diagnosis of filarial infections is based on morphological identification of microfilariae using light microscopy and requires considerable expertise, is time-consuming, and can be subjective. Loop-mediated isothermal amplification (LAMP) has advantages over microscopy or PCR because of its operational simplicity, rapidity and versatility of readout options. LAMP assays represent a major step forward in improved filarial diagnostic tools suitable for low resource settings and field applicability. The study goal was to retrospectively evaluate the performance and suitability of the O-150, RF4, and Mp419 LAMP assays for diagnosing , and infections, respectively, in humans and vectors under experimental and natural field conditions. Surveys were conducted in four health districts of Cameroon using skin snip and thick blood film methods to detect skin () and blood ( and ) dwelling microfilaria in humans. Engorged vectors (., ., and spp.) were evaluated by LAMP. Dissected, wild-caught vectors were also analyzed. LAMP showed a prevalence of 40.4% (), 17.8% () and 36.6% () versus 20.6% (), 17.4% () and 33.8% () with microscopy. spp. were dissected for microscopy and pooled for LAMP. The O-150 LAMP assay infection rate was 4.3% versus 4.1% by microscopy. spp. were dissected and analyzed individually in the LAMP assay. The RF4 LAMP assay infection rate was 23.5% versus 3.3% with microscopy. The RF4 LAMP assay also detected parasites in spp. fed on low microfilaremic volunteers. The Mp419 LAMP assay infection rate was 0.2% for and 0.04% for , while three other species were LAMP-negative. The sensitivity, species specificity, rapidity and ease of its use of these filarial LAMP assays, and validation of their performance in the field support use as alternatives to microscopy as diagnostic and surveillance tools in global health programs aimed to eliminate onchocerciasis.
丝虫感染的传统诊断方法是基于使用光学显微镜对微丝蚴进行形态学鉴定,这需要相当专业的知识,耗时且可能存在主观性。环介导等温扩增技术(LAMP)由于操作简单、快速且读出选项多样,相比显微镜检查或聚合酶链反应(PCR)具有优势。LAMP检测代表了适用于资源匮乏环境和现场应用的改进丝虫诊断工具向前迈出的重要一步。该研究的目标是回顾性评估O - 150、RF4和Mp419 LAMP检测分别在实验和自然现场条件下对人类和媒介中盘尾丝虫、罗阿丝虫和曼氏丝虫感染的诊断性能和适用性。在喀麦隆的四个卫生区进行了调查,使用皮肤切片和厚血膜方法检测人类皮肤(盘尾丝虫)和血液(罗阿丝虫和曼氏丝虫)中寄生的微丝蚴。对饱血的媒介(如蚋、库蠓和按蚊属)进行LAMP检测评估。还对野外捕获并解剖的媒介进行了分析。LAMP检测显示盘尾丝虫、罗阿丝虫和曼氏丝虫的感染率分别为40.4%、17.8%和36.6%,而显微镜检查的感染率分别为20.6%、17.4%和33.8%。对按蚊属进行解剖用于显微镜检查,并合并用于LAMP检测。O - 150 LAMP检测的感染率为4.3%,而显微镜检查为4.1%。对库蠓属进行解剖并在LAMP检测中单独分析。RF4 LAMP检测的感染率为23.5%,而显微镜检查为3.3%。RF4 LAMP检测还在吸食微丝蚴血症水平低的志愿者血液的库蠓属中检测到了寄生虫。Mp419 LAMP检测对链尾唇棘线虫的感染率为0.2%,对常现唇棘线虫的感染率为0.04%,而其他三个物种的LAMP检测结果为阴性。这些丝虫LAMP检测的敏感性、物种特异性、快速性和易用性,以及它们在现场性能的验证,支持将其用作显微镜检查的替代方法,作为全球卫生项目中旨在消除盘尾丝虫病的诊断和监测工具。