应用聚合酶链反应检测淋病奈瑟氏菌

DETECTION OF NEISSERIA GONORRHOEAE USING POLYMERASE CHAIN REACTION

根据已知淋球菌隐蔽性质粒的ｃｐｐＢ基因序列，设计了一对聚合酶链反应引物，对２１株淋球菌进行扩增均获得３９０ｂｐ片段，６株其它奈瑟氏菌及９株共生菌未能扩增出任何片段。该３９０ｂｐ片段能被限制性内切酶ＭｓｐＩ降解为２５０ｂｐ，１４０ｂｐ两个片段。经过３５个ＰＣＲ循环可检测出４００ｃｆｕ淋球菌。用ＰＣＲ法检测８２份临床标本，其敏感度、特异度、阳性预测值、阴性预测值分别为：１００％，９０％，９１．３％及１００％。

One primer set was designed according to the published complete nucleotic sequence of 4.2kb cryptic plasmid protein B of Neisseria gonorrhoeae. An amplified product of 390 bpwas obtained in 21 strains of N. gonorrhoeae,The 390 bp fragment could be digested to 250bp and 140 bp fragments by the restriction enzyme Msp I. There were no amplified productsobserved with 6 strains of non-gonococcal Neisseia and 9 genital commensals,After 35 cy-cles of amplification with the primer sets,PCR assay demonstrated a sensitivity of 400 cfu ofN. gonorrhoeae, of 82 clinical swabs,42 were positive both in culture and PCR assay,4 alsowere positive by PCR,butanegative in culture, The sensitivity,specificity,positive predictivevalue,and negative predictive value of PCR were 100%, 90%,91.3%,and 100%respective-ly.