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首页> 外文期刊>Sexually Transmitted Infections >Neisseria gonorrhoeae multi-antigen sequence typing using non-cultured clinical specimens.
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Neisseria gonorrhoeae multi-antigen sequence typing using non-cultured clinical specimens.

机译：使用非培养的临床标本进行淋病奈瑟菌多抗原序列分型。

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OBJECTIVES: The Neisseria gonorrhoeae multi-antigen sequence typing (NG-MAST) system, based on PCR amplification and sequence analysis of the gonococcal porB and tbpB genes, is widely used for molecular typing of gonococcal isolates but is not validated for non-cultured clinical samples. This study sought to examine the performance of the NG-MAST system on a range of non-cultured samples. METHODS: Nucleic acid extracts of 73 N gonorrhoeae-positive samples, comprising eight cervical swabs, nine urethral swabs, 35 urine samples, one vaginal swab, 13 rectal swabs and seven throat swabs, were analysed by NG-MAST. For 27 specimens, corresponding gonococcal isolates were also analysed and the results compared. A panel of 44 non-gonococcal Neisseria strains and 100 N gonorrhoeae-negative clinical samples were used to investigate further the specificity of the NG-MAST PCR reactions. RESULTS: PCR amplification and DNA sequencing of gonococcal porB and tbpB genes was successful for all N gonorrhoeae-positive urogenital specimens, 11 of 13 rectal swabs and four of seven throat swabs. For the 27 N gonorrhoeae-positive specimens with corresponding gonococcal isolates, the porB and tbpB sequences obtained from the non-cultured specimen were identical to those obtained from the isolate. Cross-reaction with non-gonococcal Neisseria species was observed for both the porB and tbpB PCR reactions, and proved to be problematical for NG-MAST typing of throat swab specimens. CONCLUSIONS: The NG-MAST system can successfully be applied directly to non-cultured urogenital samples, but is less suitable for extragenital specimens, particularly throat swabs, due to cross-reaction with commensal Neisseria species.
机译：目的：基于淋病球菌porB和tbpB基因的PCR扩增和序列分析，淋病奈瑟氏球菌多抗原序列分型（NG-MAST）系统被广泛用于淋球菌分离株的分子分型，但未经非临床临床验证样品。这项研究试图检查NG-MAST系统在一系列非培养样品上的性能。方法：采用NG-MAST法分析了73份淋病奈瑟氏菌阳性样本的核酸提取物，包括8个宫颈拭子，9个尿道拭子，35个尿液样本，1个阴道拭子，13个直肠拭子和7个咽喉拭子。对于27个标本，还分析了相应的淋球菌分离株并比较了结果。使用一组44个非淋球菌奈瑟菌菌株和100 N淋病奈瑟氏阴性临床样品进一步研究NG-MAST PCR反应的特异性。结果：对所有淋病奈瑟氏球菌阳性泌尿生殖道标本，13个直肠拭子中的11个和7个咽喉拭子中的4个，淋球菌porB和tbpB基因的PCR扩增和DNA测序均成功。对于具有相应淋球菌分离株的27 N淋病奈瑟氏球菌阳性标本，从非培养标本获得的porB和tbpB序列与从分离株获得的porB和tbpB序列相同。在porB和tbpB PCR反应中均观察到与非淋球菌奈瑟菌属的交叉反应，证明对NG-MAST分型的咽拭子标本是有问题的。结论：NG-MAST系统可以成功地直接应用于非培养的泌尿生殖器样本，但是由于与共生奈瑟氏球菌的交叉反应，因此不适用于生殖器外样本，尤其是咽拭子。

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