CAJ | 학술논문

目的：建立念珠状链杆菌的实时荧光定量PCR检测方法，实现该菌在多种实验动物中的快速检测。方法根据NCBI公布的念珠状链杆菌序列，设计特异性引物和探针，通过对24株标准参考菌株的扩增，验证其特异性、敏感性和重复性。运用所建立的方法对小鼠、大鼠、豚鼠、地鼠、兔、长爪沙鼠和树鼩的共823份呼吸道样本进行检测。结果用所建Taqman MGB荧光定量PCR方法在小鼠、大鼠、豚鼠、地鼠和兔中未检出念珠状链杆菌；检出普通级长爪沙鼠和树鼩中念珠状链杆菌的阳性率分别为1．5％（1／65）和61．7％（37／60）。结论本研究所建立的念珠状链杆菌Taqman MGB荧光定量PCR检测方法，能够对实验动物中念珠状链杆菌进行快速准确的检测，敏感性优于国标方法，为实验动物质量检测体系的完善奠定了基础。
목적：건립념주상련간균적실시형광정량PCR검측방법，실현해균재다충실험동물중적쾌속검측。방법근거NCBI공포적념주상련간균서렬，설계특이성인물화탐침，통과대24주표준삼고균주적확증，험증기특이성、민감성화중복성。운용소건립적방법대소서、대서、돈서、지서、토、장조사서화수구적공823빈호흡도양본진행검측。결과용소건Taqman MGB형광정량PCR방법재소서、대서、돈서、지서화토중미검출념주상련간균；검출보통급장조사서화수구중념주상련간균적양성솔분별위1．5％（1／65）화61．7％（37／60）。결론본연구소건립적념주상련간균Taqman MGB형광정량PCR검측방법，능구대실험동물중념주상련간균진행쾌속준학적검측，민감성우우국표방법，위실험동물질량검측체계적완선전정료기출。
Objective To establish a real-time quantitative PCR ( qPCR) method for detection of Streptobacillus moniliformis, which can be used to rapidly detect this pathogen in laboratory animals .Method According to the S. moniliformis sequences published in NCBI , we designed specific primers and MGB probe .The specificity, sensitivity and stability of this method were evaluated using 24 standard reference strains .Total of 823 respiratory specimens of animals including mice, rats, guinea pigs, hamsters, rabbits, Mongolian gerbils and tree shrews , were detected by this established Taqman MGB qPCR method .Results We had successfully established the S.moniliformis Taqman MGB qPCR method . S.moniliformis was not detected in the samples of mice , rats, guinea pigs, hamsters and rabbits.The positive rate of S. moniliformis was 1.5% ( 1/65 ) and 61.7% ( 37/60 ) in conventional Mongolian Gerbils and tree shrews , respectively . Conclusions Our developed qPCR method can be used to effectively detect S.moniliformis in laboratory animals .Moreover , its accuracy and sensitivity are better than the national standard method .This study laid the foundations for optimizing the quality inspection system of laboratory animals .