Instituto de Biología Molecular y Biotecnología, Carrera de Biología. Universidad Mayor de San Andrés, Calle 27 s/n, Cota Cota, La Paz, Bolivia.
Instituto de Biología Molecular y Biotecnología, Carrera de Biología. Universidad Mayor de San Andrés, Calle 27 s/n, Cota Cota, La Paz, Bolivia; Institute of Biomedicine, Department of Infectious Diseases, University of Gothenburg, Guldhedsgatan 10A, Box 480 40530 Gothenburg, Sweden.
J Virol Methods. 2022 Mar;301:114453. doi: 10.1016/j.jviromet.2021.114453. Epub 2022 Jan 3.
Group A rotavirus (RVA) is a prevalent pathogen causing acute gastroenteritis (AGE) in young children and animals. We developed an in-house ELISA (ROTA-GeFeK) for RVA detection, based on the expression of native recombinant VP6 protein in E. coli. To detect the RVA antigen, rabbit polyclonal IgG antibodies, produced against rVP6,were used as capture and detector antibodies in a sandwich ELISA. To validate the ROTA-GeFeK, 252 stool samples from children with AGE, were evaluated by conventional RT-PCR and commercial ELISA. Compared to RT-PCR, the ROTAGeFeK had a sensitivity of 88.2 % and a specificity of 94.4 %. Total detection rates with the ROTA-GeFeK, commercial ELISA and RT-PCR were 58 %, 58 % and 64 % respectively. The limit of detection was equal to 2.1 × 10 4 CCID 50 of the RVA strain RIX4414. No cross-reactivity with other enteric pathogens was observed. The RVApositive samples detected by the assay belonged to a diversity of G and [P] genotypes.This assay displayed reactivity and was proved to be useful for the detection of RVA in diarrheal samples of domestic South American Camelids. We suggest that the ROTAGeFeK can be used as an epidemiologic tool for rotavirus surveillance and for RVA detection in other animal species.
A 组轮状病毒（RVA）是一种流行的病原体，可导致幼儿和动物的急性肠胃炎（AGE）。我们基于大肠杆菌中天然重组 VP6 蛋白的表达，开发了一种用于 RVA 检测的内部 ELISA（ROTA-GeFeK）。为了检测 RVA 抗原，我们使用针对 rVP6 产生的兔多克隆 IgG 抗体作为夹心 ELISA 的捕获和检测抗体。为了验证 ROTA-GeFeK，我们用常规 RT-PCR 和商业 ELISA 评估了 252 份来自 AGE 儿童的粪便样本。与 RT-PCR 相比，ROTA-GeFeK 的灵敏度为 88.2%，特异性为 94.4%。ROTA-GeFeK、商业 ELISA 和 RT-PCR 的总检测率分别为 58%、58%和 64%。检测限等于 RVA 株 RIX4414 的 2.1×10^4CCID50。与其他肠道病原体无交叉反应。该测定法检测到的 RVA 阳性样本属于多种 G 和 [P]基因型。该检测方法显示出反应性，并被证明可用于检测南美驼属动物腹泻样本中的 RVA。我们建议 ROTA-GeFeK 可用于轮状病毒监测的流行病学工具，以及其他动物物种中 RVA 的检测。
