Sukhumsirichart W, Kiatpathomchai W, Wongteerasupaya C, Withyachumnarnkul B, Flegel T W, Boonseang V, Panyim S
Department of Biochemistry, Faculty of Medicine, Srinakharinwirot University, Sukhumvit 23, Bangkok, 10110, Thailand.
Mol Cell Probes. 2002 Dec;16(6):409-13. doi: 10.1006/mcpr.2002.0435.
A rapid and sensitive PCR-ELISA has been developed for detection of hepatopancreatic parvovirus (HPV) in Penaeus monodon. The specific primer set amplified 156 bp fragment and could detect as a little as 0.01 fg of purified HPV DNA which equivalent to three viral particles. No cross-reactivity was observed when nucleic acid templates from white spot syndrome virus, yellow-head virus, monodon baculovirus and shrimp were tested. The crude DNA simple prepared from hepatopancreas can be used as DNA template and provide a favorable result. Using this technique for detection of HPV infection in 87 carrier shrimps revealed the higher sensitivity and efficiency of detection when compared to histological examination and conventional PCR. Sixty-two percent infection was detected by PCR-ELISA from samples with HPV negative diagnosed by histological examination. Therefore, this sensitive and specific method is promisingly useful for early detection of HPV infection in broodstock, carriers and for ex situ application where large numbers of samples can be analyzed simultaneously.
已开发出一种快速灵敏的PCR-ELISA方法用于检测斑节对虾中的肝胰腺细小病毒（HPV）。特异性引物组扩增出156 bp的片段，能够检测低至0.01 fg的纯化HPV DNA，相当于三个病毒粒子。当检测来自白斑综合征病毒、黄头病毒、对虾杆状病毒和对虾的核酸模板时，未观察到交叉反应。从肝胰腺制备的粗DNA模板可用于检测，并能得到良好结果。与组织学检查和传统PCR相比，使用该技术检测87只携带HPV的对虾，显示出更高的检测灵敏度和效率。通过PCR-ELISA从组织学检查诊断为HPV阴性的样本中检测到62%的感染率。因此，这种灵敏且特异的方法有望用于亲虾、带毒虾中HPV感染的早期检测，以及可同时分析大量样本的异地应用。
