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首页> 外文期刊>Aquaculture International >A simple PCR procedure to detect white spot syndrome virus (WSSV) of shrimp, Penaeus monodon (Fabricious).
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A simple PCR procedure to detect white spot syndrome virus (WSSV) of shrimp, Penaeus monodon (Fabricious).

机译：一种简单的PCR程序，用于检测虾对虾（对虾）（Fabricious）的白斑综合症病毒（WSSV）。

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摘要

A non-stop, single tube and semi-nested polymerase chain reaction (PCR) technique with simple procedure was developed for simultaneous detection and grading the level of white spot syndrome infection in penaeid shrimp, Penaeus monodon. In this PCR procedure, three sense primers and one antisense primer with uniform annealing temperature of 55 degrees C were used. These primers amplify three PCR products (500, 300 and 200 base pairs [bp]) depending upon the severity of infection. Quantities of WSSV-DNA at 10 pg and greater gave three PCR products of 500, 300, 200 bp. A moderate concentration of WSSV-DNA, around 100 fg, gave two products of 300 and 200 bp and a low concentration of 1 fg or more gave only one PCR product of 200 bp. This PCR technique was assessed for early detection of WSSV in shrimp. In time-course infectivity experiments conducted on shrimp with WSSV, one PCR product (200 bp) was seen in hemolymph, tail tissue and gill at 3 h post infection (p.i.); two PCR products (300 and 200 bp) were seen in tail tissue, hemolymph, heart tissue and gill at 18 h p.i. At 30 h p.i., three PCR products (500, 300, 200 bp) were seen in all the organs tested. The samples collected from control animals showed negative for WSSV..
机译：开发了一种不间断的单管半巢式聚合酶链反应（PCR）技术，该技术具有简单的步骤，可同时检测对虾，对虾对虾的白斑综合症感染水平并对其进行分级。在此PCR程序中，使用了3个正义引物和1个反义引物，退火温度均匀地保持在55摄氏度。这些引物根据感染的严重程度扩增三种PCR产物（500、300和200个碱基对[bp]）。 WSSV-DNA的量为10 pg或更大时，可得到500、300、200 bp的三种PCR产物。中等浓度的WSSV-DNA，约100 fg，可得到300和200 bp的两种产物，低浓度的1 fg或更高浓度，只能得到一种200 bp的PCR产物。评估了该PCR技术以早期检测虾中的WSSV。在用WSSV对虾进行的时程感染性实验中，感染后3 h在淋巴，尾巴组织和g中发现了一种PCR产物（200 bp）（p.i。）。在18 h p.i的尾巴组织，淋巴，心脏组织和and中观察到两种PCR产物（300和200 bp）。在p.i. 30 h时，在所有测试器官中均观察到三种PCR产物（500、300、200 bp）。从对照动物收集的样品显示WSSV阴性。

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