三种方法在检测泌尿生殖道淋病奈瑟菌感染中的比较研究

Comparative study of three methods for detecting Neisseria gonorrhoeae infection in urogenital tract

目的比较反向线点杂交技术(RLB)、实时荧光PCR(FQ-PCR)及培养法在检测泌尿生殖道淋病奈瑟菌(Neisseria gonorrhoeae,NG)感染中的应用情况。方法同时采集158例患者2份泌尿生殖道标本,提取其中1份标本NG-DNA,PCR扩增NG 16SrRNA基因,然后与固定在尼龙膜上的特异性寡核苷核探针反向线点杂交;同时运用FQ-PCR检测NG的感染情况;另1份标本及时进行NG巧克力培养。结果 PCR扩增NG 16SrRNA基因的片段长度为412bp,具有较好的特异性,RLB检测158例标本中NG阳性为45例,阳性率为28.48%;FQ-PCR检测NG阳性为50例,阳性率为31.65%,二者比较差异无统计学意义(P>0.05);而培养法检测NG阳性为25例,阳性率为15.82%,明显低于RLB和FQ-PCR(P<0.01)。结论与培养法相比,RLB与FQ-PCR在检测NG方面具有简便、快速且敏感性高等优点,且二者无明显差别。

Objective To compare the application of reverse line blot hybridization assay （RLB）,real-time fluores- cent PCR（FQ-PCR） and cultivation methods in detecting Neisseria gonorrhoeae（NG） infection in urogenital tract. Methods Two urogenital specimens of 158 patients were collected at the same time, from which NG-DNA and PCR amplification NG 16s rRNA genes of one of the specimens were extracted to hybridize with the reverse line blot of specific oligonucleotide probe fixed on nylon membrane.FQ-PCR was used to detect NG infection at the same time.NG chocolate culture was performed in the other specimen.Results Fragment length of PCR amplification NG 168 rRNA gene was 412bp with good specificity.RLB detection showed that 45 out of 158 cases had positive NG.The positive rate was 28.48 %.Under the detection of FQ-PCR, NG of 50 cases were positive with a positive rate of 31.65 %. There was no significant difference between two methods（ P 〉0.05）.However,chocolate culture detection showed that 25 cases got positive NG with a positive rate of 15.82 %, significantly lower than those of RLB and FQ-PCR （ P 〈0.01）.Conclusion Compared with chocolate culture, the RLB and FQ-PCR are simple,rapid and highly sensitive and there is no significant difference between the two methods.