CAJ | 학술논문

为建立对虾无包涵体杆状病毒的快速诊断技术，应用PCR技术分别对暴发性流行病发生前、流行中和发病后的对虾样品进行了检测．根据已克隆的对虾杆状病毒部分基因组序列而设计的引物能特异性地扩增出靶DNA片段，最低可以检测1Pg的病素DNA．由典型发病症状对虾的胃、腮、肝胰腺、中肠、游泳足、肌内和心脏器官和组织中成功地检测到病毒．不同组织和器官经扩增得到的信号强弱不同：病虾的胃扩增得到的信号最强，中肠、肌肉、心脏和游泳足次之，腮和肝胰腺信号最弱，说明病毒在不同组织和器官中数量及感染程度不同．在对虾发病前及发病后，用二次PCR还能检测表面不发病呈隐性感染状态的对虾携带的病毒；而对野生健康虾的检测结果为阴性．研究表明，PCR是检测对虾暴发性流行病快速、灵敏而准确的方法，对病毒病的早期诊断、防治和高健康对虾品种的选育具有指导意义．
위건립대하무포함체간상병독적쾌속진단기술，응용PCR기술분별대폭발성류행병발생전、류행중화발병후적대하양품진행료검측．근거이극륭적대하간상병독부분기인조서렬이설계적인물능특이성지확증출파DNA편단，최저가이검측1Pg적병소DNA．유전형발병증상대하적위、시、간이선、중장、유영족、기내화심장기관화조직중성공지검측도병독．불동조직화기관경확증득도적신호강약불동：병하적위확증득도적신호최강，중장、기육、심장화유영족차지，시화간이선신호최약，설명병독재불동조직화기관중수량급감염정도불동．재대하발병전급발병후，용이차PCR환능검측표면불발병정은성감염상태적대하휴대적병독；이대야생건강하적검측결과위음성．연구표명，PCR시검측대하폭발성류행병쾌속、령민이준학적방법，대병독병적조기진단、방치화고건강대하품충적선육구유지도의의．
Specimen of Perseus chinensis were collected from hatchery ponds at Jimo, Qingdao before, during and after explcsive epidemic disease respectively in July to September 1997. In order to establish rapid dlagnsis techniques for shrimp explosive epidemic virus, diseased samples are detected by polymemse chain reaction (PCR). The pfiraers which are synthesized acccxding to the genome sequence of partially cloned shrimp haculovirus can specifically amplify the target becdloviral DNA, the viral DNA can be detected as low in quantity as I pg. The virus can be detected from the stomach, gill, halaatopanerease, min-gut, pereopod, muscle and heart of the penaeid shrimps with typical symptom of explosive epidemic disease. The positive signals vary strength from the strongest in stomach, the min-gut, muscle, heart and pereopod in the middle, to the weadest in gill and hapatopancrease. This result demonstrates the different numbers and affection extent of virus in different tissues or organs. Virus can also be detected by neeted PCR during latent and convalescent periods from shrimps showing no obvious sympton of?acute disease, whereas the revaflts on healthy wild P. Chinensis are negative. With its sensitivity, accuracy and specificity, PCR is significant for the early diagnosis of viral diaease and the high healthy stock brdlng of penaeid shrimp.