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Detection of Dual-Infected Cases of Adenoviruses and Coxsackieviruses Type B by Real-Time PCR but not by the Conventional Viral Culture Technique

机译:通过实时荧光定量PCR(而非常规病毒培养技术)检测B型腺病毒和柯萨奇病毒双重感染病例

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The aim of this study was to evaluate the applicability of diagnostic methods for dual-infected cases of human adenoviruses (AdVs) and coxsackieviruses type B (CBs). For this purpose, 100 nasopharyngeal samples from patients with acute exudative tonsillitis and clinically suspected AdV infection were analyzed. Using PCR and real-time PCR techniques for AdVs and CBs, we found 86 AdVs-only positive samples; we also found five dual-infected samples containing 5.4 × 10~5 to 7.1 × 10~8 copies/mL of AdV genomes and 1.4 × 10~4 to 1.3 ×10~9 copies/mL of CB genomes. By viral culture using A549 cells, two co-infected samples, which contained over 10~8 copies/mL of AdV genomes and < 10~5 copies/mL of CB genomes, became AdV dominant, while three samples with less than 2.0 × 10~6 copies/mL of AdV genomes became CB dominant. An immunochromatography kit for diagnosing AdVs at the bedside was positive for 3/5 dual-infected patients, and PCR techniques for AdVs and CBs were both positive for 5/5. Viral culture is usually considered to be the gold standard for AdV diagnosis, but our results demonstrate the importance of PCR applications for the detection of AdV and CB genomes, particularly in clinical cases of suspected AdV infection. Even though the sample size of dual infection (n=5) is small, our results show the existence of dual infection cases which were difficult to diagnose by viral culture alone.
机译:这项研究的目的是评估诊断方法对人腺病毒(AdVs)和B型柯萨奇病毒(CBs)双重感染病例的适用性。为此,分析了急性渗出性扁桃体炎和临床疑似AdV感染患者的100份鼻咽样本。使用针对AdV和CB的PCR和实时PCR技术,我们发现了86个仅AdVs的阳性样本。我们还发现了五个双重感染的样本,分别含有5.4×10〜5至7.1×10〜8拷贝/ mL的AdV基因组和1.4×10〜4至1.3×10〜9拷贝/ mL的CB基因组。通过使用A549细胞进行病毒培养,两个共感染的样本包含AdV基因组超过10〜8拷贝/ mL,CB基因组<10〜5拷贝/ mL成为AdV优势,而三个样本的2.0×10以下大约6拷贝/ mL的AdV基因组成为CB优势。用于床旁AdVs诊断的免疫色谱试剂盒对3/5双重感染患者呈阳性反应,而对AdVs和CBs的PCR技术对5/5均为阳性。病毒培养通常被认为是诊断AdV的金标准,但我们的结果证明PCR应用对于检测AdV和CB基因组的重要性,尤其是在怀疑有AdV感染的临床病例中。即使双重感染的样本量很小(n = 5),我们的结果也表明存在双重感染病例,仅通过病毒培养难以诊断。

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