Multiplex PCR methods for detection of several viruses associated with canine respiratory and enteric diseases
Multiplex PCR methods for detection of several viruses associated with canine respiratory and enteric diseases
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用于检测与犬呼吸道和肠道疾病相关的多种病毒的多重 PCR 方法
DOI：
10.1371/journal.pone.0213295
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发表时间：
2019-03-04
期刊：
影响因子：
3.7
通讯作者：
Li, Shoujun
中科院分区：
文献类型：
--
作者：
Hao, Xiangqi;Liu, Ruohan;Li, Shoujun
病毒性呼吸道和肠道感染是犬病毒性疾病最常见的原因。多种病原体的感染在许多情况下发生。快速诊断这些多重感染对于提供及时有效的治疗非常重要。为了提高诊断水平，本研究开发了两种新的多重聚合酶链反应（mPCR），使用两种不同的引物混合物同时检测犬呼吸道病毒（CRV）和犬肠道病毒（CEV）。这些病毒包括犬腺病毒2型（CAV-2）、犬瘟热病毒（CDV）、犬流感病毒（CIV）、犬副流感病毒（CPIV）、犬圆环病毒（CanineCV）、犬冠状病毒（CCoV）和犬细小病毒（CPV）。结果表明，两种方法的检测限均为1 × 104拷贝。使用新型mPCR方法作为临床试验，评价了从具有呼吸道疾病或肠道疾病症状的犬中采集的20份鼻拭子（NS）和20份肛门拭子（AS）。当应用于这些呼吸道样本和肠道样本时，mPCR方案可以同时检测7种病毒，从而可以快速调查CRV（CAV-2，CDV，CIV和CPIV）和CEV（CAV-2，CanineCV，CCoV和CPV）的状态，并及时评估合并感染。我们的研究为犬的快速鉴别诊断和流行病学监测提供了一个有效和准确的工具。
Viral respiratory and intestinal infections are the most common causes of canine viral illness. Infection with multiple pathogens occurs in many cases. Rapid diagnosis of these multiple infections is important for providing timely and effective treatment. To improve diagnosis, in this study, two new multiplex polymerase chain reactions (mPCRs) were developed for simultaneous detection of canine respiratory viruses (CRV) and canine enteric viruses (CEV) using two separate primer mixes. The viruses included canine adenovirus type 2 (CAV-2), canine distemper virus (CDV), canine influenza virus (CIV), canine parainfluenza virus (CPIV), canine circovirus (CanineCV), canine coronavirus (CCoV) and canine parvovirus (CPV). The sensitivity of the mPCR results showed that the detection limit of both mPCR methods was 1×104 viral copies. Twenty nasal swabs (NS) and 20 anal swabs (AS) collected from dogs with symptoms of respiratory disease or enteric disease were evaluated using the novel mPCR methods as a clinical test. The mPCR protocols, when applied to these respiratory specimens and intestinal samples, could detect 7 viruses simultaneously, allowing rapid investigation of CRV (CAV-2, CDV, CIV and CPIV) and CEV (CAV-2, CanineCV, CCoV and CPV) status and prompt evaluation of coinfection. Our study provides an effective and accurate tool for rapid differential diagnosis and epidemiological surveillance in dogs.