CAJ | 학술논문

目的建立同时检测诺如病毒、轮状病毒、星状病毒和甲肝病毒多重RT-PCR检测方法.方法以四种食源性病毒的高度保守区为靶序列设计特异引物,优化反应体系和条件,确定多重RT-PCR检测四种病毒的特异性和灵敏度,并初步应用于临床样本中四种病毒的同时检测.结果在灵敏度试验中得到的轮状病毒、诺如病毒和星状病毒稳定的最高检测限均为50 pg/ml,甲肝病毒为100 pg/ml.在128份临床粪便样本中,其中轮状病毒阳性62份(48.44%),诺如病毒阳性8份(6.25%),星状病毒阳性11份(8.59%),甲肝病毒阳性4份(3.12%).结论研究所建立的多重RT-PCR方法,在实际应用中能同时处理大量的样本,提高PCR检测方法的能力,可以应用于临床病例的诊断和流行病学调查等研究.
목적 건립동시검측낙여병독、륜상병독、성상병독화갑간병독다중RT-PCR검측방법.방법 이사충식원성병독적고도보수구위파서렬설계특이인물,우화반응체계화조건,학정다중RT-PCR검측사충병독적특이성화령민도,병초보응용우림상양본중사충병독적동시검측.결과 재령민도시험중득도적륜상병독、낙여병독화성상병독은정적최고검측한균위50 pg/ml,갑간병독위100 pg/ml.재128빈림상분편양본중,기중륜상병독양성62빈(48.44%),낙여병독양성8빈(6.25%),성상병독양성11빈(8.59%),갑간병독양성4빈(3.12%).결론 연구소건립적다중RT-PCR방법,재실제응용중능동시처리대량적양본,제고PCR검측방법적능력,가이응용우림상병례적진단화류행병학조사등연구.
Objective To establish a method for simultaneous detection of norovirus (NV),rotavirus (RV), astrovirus (AV) and hepatitis A virus (HAV) by multiplex reverse transcriptionpolymerase chain reaction (RT-PCR). Methods Specific primers of the four viruses were designed based on the high conserved sequences, the reaction system and conditions optimized and the specificity and sensitivity confirmed. The method was then applied to detect the four viruses in clinical samples. Results The steady detection limits were 100 pg/ml for hepatitis A virus, 50 pg/ml for rotavirus, norovirus and astrovirus respectively. When the developed method was used to detect clinical fecal samples, 62(48.44%)were iden tified as rotavirus, 8 (6.25%) as norovirus, 11 (8.59%) as astrovirus and 4 (3.12%) as hepatitis A virus in a total of 128 samples. Conclusion Data from our study showed that multiplex RTPCR system could be used to simultaneously detect the four viruses in routine monitoring and risk assessment in disease outbreaks with high specificity and sensitivity.