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作者设计并合成了钩端螺旋体(简称钩体)犬群大型Moulton株23S rRNA基因的一对引物,即引物A_1 5’GAT CTA ATT CGC TGT AGC AGG~3’及引物B_1 5’ACT TTC ACCCTC TAT GGT CGG~3’用于聚合酶链反应(PCR)检测不同群型的问号状钩体,结果该引物不能使双曲钩体、细螺旋体和其它致病微生物及人白细胞DNA等扩增特异性片段,并用该对引物扩增早期钩体病患者和正常人及其它疾病患者的血清标本(以临床确诊、血培养及MAT阳性为金标准),检测结果表明:PCR诊断钩体病的敏感性为92.00％,特异性为94.35％,准确性为92.54％,阳性预测值为98.17％,阴性预测值为78.13％,阳性拟然比为16.25,阴性拟然比为0.0848。由此表明PCR扩增钩体235 rRNA基因,是早期钩体病诊断的一种敏感、特异、快速而简便的方法。
The authors designed and synthesized a pair of primers for 23S rRNA gene of Leptospira interrogans large Moulton strain, that is, primer A_1 5’GAT CTA ATT CGC TGT AGC AGG ~ 3 ’and primer B_1 5’ACT TTC ACCCTC TAT GGT CGG ~ 3 ’was used for the polymerase chain reaction (PCR) detection of different groups of tick-like leptospira. As a result, the primer could not amplify the DNA of Leptospira, leptospira and other pathogenic microorganisms and human leukocyte The primers were used to amplify the serum samples of patients with early leptospirosis and normal and other diseases (clinically confirmed, blood culture and MAT positive). The results showed that PCR could be used to diagnose the leptospirosis susceptibility 92.00%, specificity was 94.35%, accuracy was 92.54%, positive predictive value was 98.17%, negative predictive value was 78.13%, positive predictive ratio was 16.25, negative predictive value was 0.0848. Thus, it is indicated that PCR amplification of 235 rRNA gene of leptospira is a sensitive, specific, rapid and simple method for the diagnosis of early leptospirosis.