Rajan J Joseph Sahaya, Praveena P Ezhil, Bhuvaneswari T, Jithendran K P
Aquatic Animal Health and Environment Division, Central Institute of Brackishwater Aquaculture, 75, Santhome High Road, Chennai, Tamil Nadu 600 028 India.
Virusdisease. 2016 Jun;27(2):123-9. doi: 10.1007/s13337-016-0313-0. Epub 2016 Apr 29.
Viral encephalopathy and retinopathy otherwise known as viral nervous necrosis (VNN) is a neuropathological condition affecting more than 50 fish species worldwide, mostly marine. Different PCR protocols with specific primers were reported from many countries for confirmation of VNN in fishes. In the present study, two pairs of primers were designed and evaluated for the diagnosis of clinical and subclinical cases of infections from field. These primers designated as BARL-F1/BARL-R1 amplified a 902 bp product in the variable region (T4) of the coat protein gene by first step PCR. Nested PCR primers BARL-F2/BARL-R2 amplified a fragment of 313 bp. The results were comparable with other commonly used primer sets such as F2/R3 and RG668f/RG919r primers. These new primers could detect betanodavirus in standard reference samples containing low, moderate and high viral load. Known positive and negative control samples of fish also revealed a predictive value of 100 % by RT-PCR diagnosis.
病毒性脑病和视网膜病，也被称为病毒性神经坏死病（VNN），是一种神经病理学疾病，影响着全球50多种鱼类，其中大多数为海洋鱼类。许多国家都报道了使用特定引物的不同PCR检测方法，用于确诊鱼类的VNN。在本研究中，设计并评估了两对引物，用于诊断野外感染的临床和亚临床病例。这些引物命名为BARL-F1/BARL-R1，通过第一步PCR在衣壳蛋白基因的可变区（T4）扩增出一个902 bp的产物。巢式PCR引物BARL-F2/BARL-R2扩增出一个313 bp的片段。结果与其他常用引物组如F2/R3和RG668f/RG919r引物相当。这些新引物能够在含有低、中、高病毒载量的标准参考样本中检测到β-诺达病毒。已知的鱼类阳性和阴性对照样本通过RT-PCR诊断也显示出100%的预测价值。
