评价环介导等温扩增检测试剂盒和抗原酶联免疫吸附试验在孟加拉国消除后内脏利什曼病的检测和管理中的应用。

Evaluation of Loopamp™ Detection Kit and Antigen ELISA for Post-Elimination Detection and Management of Visceral Leishmaniasis in Bangladesh.

机构信息

Emerging infections and Parasitology laboratory, Nutrition and Clinical Service Division, International Centre for Diarrhoeal Disease Research, Bangladesh, Dhaka, Bangladesh.

Neglected Tropical Diseases, Foundation for Innovative New Diagnostics, Geneva, Switzerland.

出版信息

Front Cell Infect Microbiol. 2021 Apr 26;11:670759. doi: 10.3389/fcimb.2021.670759. eCollection 2021.

DOI:10.3389/fcimb.2021.670759

PMID:33981632

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8108992/

Abstract

With reduced prevalence of visceral leishmaniasis (VL) in the Indian subcontinent (ISC), direct and field deployable diagnostic tests are needed to implement an effective diagnostic and surveillance algorithm for post-elimination VL control. In this regard, here we investigated the diagnostic efficacies of a loop-mediated isothermal amplification (LAMP) assay (Loopamp™ Detection Kit, Eiken Chemical CO., Ltd, Japan), a real-time quantitative PCR assay (qPCR) and the antigen ELISA (CLIN-TECH, UK) with different sampling techniques and evaluated their prospect to incorporate into post-elimination VL control strategies. Eighty clinically and rK39 rapid diagnostic test confirmed VL cases and 80 endemic healthy controls were enrolled in the study. Peripheral blood and dried blood spots (DBS) were collected from all the participants at the time of diagnosis. DNA was extracted from whole blood (WB) and DBS silica columns (QIAGEN) and boil & spin (B&S) methods and tested with qPCR and Loopamp. Urine was collected from all participants at the time of diagnosis and was directly subjected to the antigen ELISA. 41 patients were followed up and urine samples were collected at day 30 and day 180 after treatment and ELISA was performed. The sensitivities of the Loopamp-WB(B&S) and Loopamp-WB(QIA) were 96.2% (95% CI 89·43-99·22) and 95% (95% CI 87·69-98·62) respectively. The sensitivity of Loopamp-DBS(QIA) was 85% (95% CI 75·26- 92·00). The sensitivities of the qPCR-WB(QIA) and qPCR-DBS(QIA) were 93.8% (95% CI 86·01-97·94) and 72.5% (95% CI 61·38-81·90) respectively. The specificity of all molecular assays was 100%. The sensitivity and specificity of the antigen ELISA were 97.5% (95% CI 91·47-99·70) and 91.95% (95% CI 84·12-96·70) respectively. The antigen ELISA depicted clinical cure at day 180 in all the followed-up cases. Efficacy and sustainability identify the Loopamp-WB(B&S) and the antigen ELISA as promising and minimally invasive VL diagnostic tools to support VL diagnostic and surveillance activities respectively in the post-elimination era.

摘要

随着印度次大陆（ISC）内脏利什曼病（VL）发病率的降低，需要直接和现场部署的诊断检测方法来实施有效的诊断和监测消除后 VL 控制的算法。在这方面，我们在这里研究了环介导等温扩增（LAMP）检测试剂盒（Eiken Chemical CO.，Ltd.，日本）、实时定量 PCR 检测试剂盒（qPCR）和抗原 ELISA（CLIN-TECH，英国）的诊断效果，采用不同的采样技术，并评估了它们纳入消除后 VL 控制策略的前景。80 例临床和 rK39 快速诊断试验确诊的 VL 病例和 80 例地方性健康对照者纳入研究。所有参与者在诊断时均采集外周血和干血斑（DBS）。从全血（WB）和 DBS 硅柱（QIAGEN）和煮沸和旋转（B&S）方法中提取 DNA，并与 qPCR 和 Loopamp 一起进行检测。所有参与者在诊断时采集尿液，并直接进行抗原 ELISA。41 例患者接受随访，在治疗后 30 天和 180 天采集尿液样本并进行 ELISA。Loopamp-WB（B&S）和 Loopamp-WB（QIA）的灵敏度分别为 96.2%（95%CI 89·43-99·22）和 95%（95%CI 87·69-98·62）。Loopamp-DBS（QIA）的灵敏度为 85%（95%CI 75·26-92·00）。qPCR-WB（QIA）和 qPCR-DBS（QIA）的灵敏度分别为 93.8%（95%CI 86·01-97·94）和 72.5%（95%CI 61·38-81·90）。所有分子检测的特异性均为 100%。抗原 ELISA 的灵敏度和特异性分别为 97.5%（95%CI 91·47-99·70）和 91.95%（95%CI 84·12-96·70）。在所有随访病例中，抗原 ELISA 在第 180 天显示临床治愈。Loopamp-WB（B&S）和抗原 ELISA 的功效和可持续性分别确定为有前途的和微创 VL 诊断工具，以分别支持消除后的 VL 诊断和监测活动。