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Cell culture demonstrates superior sensitivity over one step real time RT PCR and nested VP1 amplification for Enteroviruses
Journal of Virological Methods ( IF 2.2 ) Pub Date : 2020-10-14 , DOI: 10.1016/j.jviromet.2020.113994
Auwal Idris Kabuga 1 , Ahmad Nejati 1 , Parastoo Soheili 1 , Soodeh Yousefipoor 1 , Maryam Yousefi 1 , Yaghoob Mollaiee 1 , Shohreh Shahmahmoodi 2
Affiliation  

This study evaluated and compared the sensitivity profile of routine cell culture, nested VP1 amplification and one step real time RT PCR for Enteroviruses. Serially diluted spiked samples of four model viruses (EV71, CVA16, CVB5 and PV1) and 32 true positive samples including Poliovirus (PV1 & PV3), Coxsackie virus (CVB5, CVB3, CVB1 & CVA4, 10, 16), Echovirus (Echo 6, 7, 11, 13, 18, 25 & 30) and Enterovirus 71 (E71), and 32 true negative stool samples were subjected to cell culture, nested RT PCR and one step real time RT PCR. The result of sensitivity test indicated superior sensitivity with one step real time RT PCR (75 %, 24/32) against cell culture (71.9 %, 23/32) and nested RT PCR (65.6 %, 21/32). The most specific test was cell culture (100 %, 32/32), followed by nested RT PCR (96.9 %, 31/32). Positive predictive values were 100 %: 23/23, 95.5 %; 21/22 and 88.9 %; 24/27, for cell culture, nested RT PCR and one step real time RT PCR, respectively, and one step real time RT PCR had the highest negative predictive value (78.4 %, 29/37). Overall result indicate relatively high analytical sensitivity with all the tests, suggesting superior performance by cell culture. Therefore, cell culture is the gold standard. However, considering intensive nature of cell cultures and prolong window for results, it is wise to consider one step real time RT PCR in routine diagnosis for its added advantages. Meanwhile, selecting a combination of tests can maximize detection, depending on the laboratory strength.



中文翻译:

细胞培养对肠道病毒的一步实时 RT PCR 和嵌套 VP1 扩增显示出更高的灵敏度

本研究评估并比较了常规细胞培养、巢式 VP1 扩增和一步法实时 RT PCR 对肠道病毒的敏感性特征。四种模型病毒(EV71、CVA16、CVB5 和 PV1)的连续稀释加标样本和 32 个真阳性样本,包括脊髓灰质炎病毒(PV1 和 PV3)、柯萨奇病毒(CVB5、CVB3、CVB1 和 CVA4、10、16)、回声病毒(Echo 6) 、7、11、13、18、25 和 30) 和肠道病毒 71 (E71) 和 32 个真阴性粪便样本进行细胞培养、巢式 RT PCR 和一步实时 RT PCR。灵敏度测试结果表明,一步实时 RT PCR (75 %, 24/32) 对细胞培养 (71.9 %, 23/32) 和巢式 RT PCR (65.6 %, 21/32) 具有优异的灵敏度。最具体的测试是细胞培养(100%,32/32),其次是巢式 RT PCR(96.9%,31/32)。阳性预测值为 100 %:23/23,95.5 %;21/22 和 88.9%;24/27,对于细胞培养,巢式 RT PCR 和一步实时 RT PCR 分别和一步实时 RT PCR 具有最高的阴性预测值 (78.4 %, 29/37)。总体结果表明,所有测试的分析灵敏度都相对较高,表明细胞培养具有优异的性能。因此,细胞培养是金标准。然而,考虑到细胞培养的密集性和延长结果窗口,在常规诊断中考虑一步实时 RT PCR 是明智的,因为它具有额外的优势。同时,根据实验室的实力,选择测试组合可以最大限度地提高检测率。一步实时 RT PCR 具有最高的阴性预测值 (78.4 %, 29/37)。总体结果表明,所有测试的分析灵敏度都相对较高,表明细胞培养具有优异的性能。因此,细胞培养是金标准。然而,考虑到细胞培养的密集性和延长结果窗口,在常规诊断中考虑一步实时 RT PCR 是明智的,因为它具有额外的优势。同时,根据实验室的实力,选择测试组合可以最大限度地提高检测率。一步实时 RT PCR 具有最高的阴性预测值 (78.4 %, 29/37)。总体结果表明,所有测试的分析灵敏度都相对较高,表明细胞培养具有优异的性能。因此,细胞培养是金标准。然而,考虑到细胞培养的密集性和延长结果窗口,在常规诊断中考虑一步实时 RT PCR 是明智的,因为它具有额外的优势。同时,根据实验室的实力,选择测试组合可以最大限度地提高检测率。考虑到细胞培养的密集性和延长结果窗口,在常规诊断中考虑一步实时 RT PCR 是明智的,因为它具有额外的优势。同时,根据实验室的实力,选择测试组合可以最大限度地提高检测率。考虑到细胞培养的密集性和延长结果窗口,在常规诊断中考虑一步实时 RT PCR 是明智的,因为它具有额外的优势。同时,根据实验室的实力,选择测试组合可以最大限度地提高检测率。

更新日期:2020-10-30
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