Guangdong Key Laboratory of Laboratory Animals, Guangdong Laboratory Animals, Monitoring Institute, Guangzhou, China.
College of Animal Science, South China Agricultural University, Guangzhou, China.
Biomed Res Int. 2018 Nov 6;2018:5035139. doi: 10.1155/2018/5035139. eCollection 2018.
Porcine deltacoronavirus (PDCoV) is a newly discovered coronavirus, which belongs to the family Coronaviridae. It causes watery diarrhea, vomiting, and dehydration in newborn piglets. A sensitive RT-PCR method is urgently required to detect PDCoV infection. In this study, we developed and evaluated a conventional RT-PCR assay and a SYBR green-based real-time RT-PCR assay that targeted the PDCoV gene. Both assays are specific and have the same limit of detection at 2 × 10 copies of RNA molecules per reaction. Eighty-four clinical samples were subjected to both conventional RT-PCR and real-time RT-PCR, and the same positive rate (41.7%) was achieved, which was much higher than the positive rate (26.2%) using a previously described one-step RT-PCR technique. In summary, a conventional RT-PCR technique was successfully established for the detection of PDCoV with the same detection limit as a SYBR green-based real-time RT-PCR assay.
猪德尔塔冠状病毒（PDCoV）是一种新发现的冠状病毒，属于冠状病毒科。它会导致新生仔猪出现水样腹泻、呕吐和脱水。目前急需一种灵敏的 RT-PCR 方法来检测 PDCoV 感染。在这项研究中，我们开发并评估了一种针对 PDCoV 基因的常规 RT-PCR 检测方法和一种 SYBR Green 实时 RT-PCR 检测方法。这两种方法均具有特异性，检测限均为每个反应 2×10 个 RNA 分子拷贝。对 84 份临床样本进行了常规 RT-PCR 和实时 RT-PCR 检测，阳性率相同（41.7%），明显高于先前描述的一步法 RT-PCR 技术（26.2%）。总之，成功建立了一种常规 RT-PCR 技术，用于检测 PDCoV，其检测限与 SYBR Green 实时 RT-PCR 检测方法相同。
