Evaluation of a commercial immunochromatographic strip assay (ICT) for rapid detection of bovine, porcine and human Rotavirus A

The Microbe Pub Date : 2024-09-28 DOI:10.1016/j.microb.2024.100182

Bilal Ahmad Malla , Zunjar Baburao Dubal , Shubham Saini , Obli Rajendran VinodhKumar , Aquil Mohmad , P.T. Waseem Mirsab , Mufeeda Beegum , Vibha Singh , Kiran Narayan Bhilegaonkar

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Abstract

The most frequent diarrhoeal pathogen “Rotavirus” is commonly detected by Ribo Nucleic Acid- Polyacrylamide Gel Electrophoresis (RNA-PAGE), Reverse Transcription Polymerase Chain Reaction (RT-PCR) and Immunoassay (Enzyme Linked Immunosorbent Assay) in humans. However, use of these tests is limited in animals therefore, penside test like Immunochromatographic test/Lateral Flow Assay (ICT/LFA) is necessary. The present study was carried out to compare the diagnostic efficacy of rapid commercial ICT strip assay, RNA-PAGE and RT-PCR for specific detection of Rotavirus A (RVA) from stool samples of calves, piglets and children. A total of 313 faecal samples were collected between November 2022 to April 2023 from children below 5years of age (n = 100), calves (n = 100) and piglets (n = 113) ≤ 3 months of age and were screened by RNA-PAGE, RT-PCR and commercial ICT kit for rotavirus A.

The overall positivity of RVA from human, calves and piglets using RNA-PAGE, RT-PCR and LFA was found to be 35 % (35/100), 8 % (8/100) and 14.16 % (16/113), respectively. Higher positivity of rotavirus was recorded in male children (44.26 %, 27/61) than in female children (20.51 %, 8/39). The kappa agreement between LFA and RT-PCR was 0.79 (substantial agreement); between LFA and RNA-PAGE was 0.61 (substantially low agreement) and between RNA-PAGE and RT-PCR was found to be 0.75 (substantial agreement). Relative diagnostic sensitivity and specificity of LFA in comparison with RT-PCR was 74.54 % and 98.44 %, respectively and in comparison with RNA-PAGE was 72.97 % and 93.47 %, respectively. RNA-PAGE in comparison with RT-PCR is having diagnostic sensitivity and specificity of 65.45 % and 99.6 %, respectively. In conclusion, the ICT strip assay is a rapid, convenient and effective method with satisfactory efficacy for detection of RVA from children, calves and piglets. ICT fulfilled the WHO ASSURED (Affordable, Sensitive, Specific, User-friendly, Rapid and robust, Equipment-free and Delivered) criteria for point-of-care testing. It can be useful in determining rotavirus outbreaks in resource-limited settings.

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评估用于快速检测牛、猪和人轮状病毒 A 的商用免疫层析技术 (ICT)

在人类中，最常见的腹泻病原体 "轮状病毒 "通常通过核酸-聚丙烯酰胺凝胶电泳（RNA-PAGE）、反转录聚合酶链式反应（RT-PCR）和免疫测定（酶联免疫吸附试验）来检测。然而，这些检测方法在动物身上的应用有限，因此有必要进行免疫层析检测/侧流检测（ICT/LFA）等笔侧检测。本研究比较了快速商用 ICT 条形检测法、RNA-PAGE 和 RT-PCR 对小牛、仔猪和儿童粪便样本中轮状病毒 A（RVA）特异性检测的诊断效果。在2022年11月至2023年4月期间，共收集了313份粪便样本，分别来自5岁以下的儿童（n = 100）、小牛（n = 100）和3个月以下的仔猪（n = 113），并通过RNA-PAGE、RT-PCR和商用ICT试剂盒进行了轮状病毒A的筛查。通过RNA-PAGE、RT-PCR和LFA检测发现，人、犊牛和仔猪的RVA总阳性率分别为35%（35/100）、8%（8/100）和14.16%（16/113）。男性儿童的轮状病毒阳性率（44.26%，27/61）高于女性儿童（20.51%，8/39）。LFA与RT-PCR之间的卡帕值为0.79（高度一致）；LFA与RNA-PAGE之间的卡帕值为0.61（高度一致）；RNA-PAGE与RT-PCR之间的卡帕值为0.75（高度一致）。与 RT-PCR 相比，LFA 的相对诊断灵敏度和特异性分别为 74.54 % 和 98.44 %；与 RNA-PAGE 相比，LFA 的相对诊断灵敏度和特异性分别为 72.97 % 和 93.47 %。与 RT-PCR 相比，RNA-PAGE 的诊断灵敏度和特异性分别为 65.45 % 和 99.6 %。总之，ICT 检测条是一种快速、方便、有效的方法，对检测儿童、犊牛和仔猪的 RVA 有令人满意的效果。ICT 符合世卫组织 ASSURED（经济实惠、灵敏、特异、使用方便、快速稳健、无需设备、可送达）标准，可用于护理点检测。在资源有限的情况下，它可用于确定轮状病毒的爆发。

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The Microbe

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