gyrA和parE基因检测在脲原体基因分型中的应用

目的 研究gyrA和parE基因检测在脲原体基因分型中的作用.方法 脲原体培养与药敏分析用Mycoplasma IST检测试剂盒;在脲原体培养阳性者中选取对喹诺酬耐药标本60份,PCR扩增gyrA和parE基因,扩增产物经测序分析后与基因库中的脲原体各血清型进行比对.结果 gyrA扩增片段在血清型1、3、6、14之间核苷酸序列相似性为100%,在血清型2,4、5、7～13之间核苷酸序列相似性100%,两组之间核苷酸序列相似性91%.parE扩增片段在血清型1、3、6、14之间的核苷酸序列相似性为98%～99%;pare扩增片段在血清犁2、5、7、8、11之间核苷酸序列相似性100%,在血清型4、12、13之间核苷酸序列相似性100%,两组之间核苷酸序列相似性为90%.60份标本中微小脲原体(Ureaplasma parvum,Up)占68.3%(41/60),解脲脲原体(Ureaplasma urealyticum,Uu)占21.7%(13/60),两型混合感染占10%(6/60).在Up中,血清型3占48.8%(20/41).结论 gyrA检测可把脲原体分为微小脲原体和解脲脲原体两个基因型,parE检测可把微小脲原体分为4个亚型,分别与血清型1、3、6、14完全一致,其中血清型3感染最常见.

Application of gyrA and pare gene detection in genotyping of Ureaplasma spp.

Objective To evaluate the contribution of gyrA and parE detection in Ureaplasma genotyping.Methods Sixty Ureaplasma isolates were selected with the Mycoplasma IST kit.The gyrA and parE were amplified by PCR.The DNA was sequenced and compared with the corresponding sequences in GenBank.Results The nucleotide sequence of gyrA had 100% identity in serovar 1,3,6,14 and 100%identity in serovar 2,4,5,7～13,too.But the sequence had 91%identity between the two groups.The nucleotide sequence of parE had 98%～99% identity in serovar 1,3,6,14.And it had 100% identity in erovar 2,5,7,8,11 and 100% identity in serovar4,12,13.But it had only 90% identity between the two groups.Ureaplasma parvum(Up),Ureaplasma urealyticum(Uu)and Up+Uu infection were found 68.3%(41/60),21.7%(13/60)and 10%(6/60) of clinical specimens,respectively.In Up isolates,serovar 3 was 48.8%(20/41).Conclusion Ureaplasma can be divided into two genotypes(Up and Uu)by gyrA analysis.And Up can be divided into four subtypes which correspond to serovar 1,3,6,14,respectively.Serovar 3 is the main isolate in our research.