Center of Pediatric and Adolescent Medicine, University Medical Center, Mainz, Germany.
Department of Pediatric and Adolescent Medicine, Bugando Medical Centre, Mwanza, Tanzania.
Malar J. 2021 Feb 1;20(1):66. doi: 10.1186/s12936-021-03595-4.
Children are the most vulnerable group affected by malaria and other tropical, vector-borne diseases in low-resource countries. Infants presenting with acute onset fever represent a major sector of outpatient care in the Lake Victoria region. Misclassification and overuse of antibiotics and anti-malarial medications are consistent problems. Identifying the prevalent mosquito-borne pathogens in the region will reduce the prescription of non-indicated medicines.
The literature was reviewed focusing on the mosquito-borne pathogens most prevalent in sub-Saharan Africa. Accordingly, an assay comprised of a multiplex-reverse transcriptase-polymerase chain reaction and an enzyme-linked immunosorbent assay (multiplex-RT-PCR-ELISA) was designed and validated in its ability to identify and differentiate nine human mosquito-borne pathogens including eight arboviruses and Plasmodium sp., the aetiologic agents of malaria. Blood samples obtained from 132 children suspected of having malaria were spotted and preserved on Whatman 903 protein sample cards. Multiplex-RT-PCR-ELISA analysis was assessed and compared to results obtained by blood smear microscopy and the malaria rapid diagnostic test (RDT).
Nine out of nine pathogens were amplified specifically by the multiplex-RT-PCR-ELISA panel. Twenty-seven out of 132 paediatric patients presenting with acute fever were infected with Plasmodium sp., confirmed by multiplex-RT-PCR. The results of blood smear microscopy were only 40% sensitive and 92.8% specific. The malaria RDT, on the other hand, detected acute Plasmodium infections with 96.3% sensitivity and 98.1% specificity. The preservation of Plasmodium sp. in clinical sera and whole blood samples spotted on sample cards was evaluated. The duration of successful, sample card storage was 186 to 312 days.
Reliable, easy-to-use point of care diagnostic tests are a powerful alternative to laboratory-dependent gold standard tests. The multiplex-RT-PCR-ELISA amplified and identified nine vector-borne pathogens including Plasmodium sp. with great accuracy. Translation of improved diagnostic approaches, i.e., multiplex-RT-PCR-ELISA, into effective treatment options promises to reduce childhood mortality and non-indicated prescriptions.
儿童是资源匮乏国家中受疟疾和其他热带病影响最严重的脆弱群体。急性发作发热的婴儿是维多利亚湖地区门诊护理的主要人群。抗生素和抗疟药物的错误分类和过度使用一直是存在的问题。确定该地区流行的蚊子传播病原体将减少非指征性药物的处方。
重点回顾了撒哈拉以南非洲地区最常见的蚊子传播病原体的文献。因此，设计并验证了一种由多重逆转录聚合酶链反应和酶联免疫吸附测定（多重 RT-PCR-ELISA）组成的检测方法，以识别和区分包括 8 种虫媒病毒和疟原虫在内的 9 种人类蚊子传播病原体，疟原虫是疟疾的病因。从 132 名疑似患有疟疾的儿童中采集血液样本，点样并保存在 Whatman 903 蛋白质样品卡上。对多重 RT-PCR-ELISA 分析进行了评估，并与血涂片显微镜检查和疟疾快速诊断检测（RDT）的结果进行了比较。
九种病原体均被多重 RT-PCR-ELISA 检测板特异性扩增。132 名急性发热的儿科患者中，有 27 人被证实感染疟原虫，通过多重 RT-PCR 检测到。血涂片显微镜检查的结果仅为 40%敏感和 92.8%特异。另一方面，疟疾 RDT 检测急性疟原虫感染的敏感性为 96.3%，特异性为 98.1%。评估了临床血清和点样在样品卡上的全血样本中疟原虫的保存情况。成功保存样本卡的时间为 186 至 312 天。
可靠、易于使用的即时检测是对依赖实验室的金标准检测的有力替代。多重 RT-PCR-ELISA 可高度准确地扩增和识别包括疟原虫在内的 9 种蚊子传播病原体。将改进的诊断方法（如多重 RT-PCR-ELISA）转化为有效的治疗方案有望降低儿童死亡率和非指征性处方。
