| 高剑云,陈建忠,赵金方,周坚红.淋病奈瑟菌pI优势基因型及其G120/A121突变与耐药性关系[J].Chinese journal of Epidemiology,2010,31(4):442-446 |
| 淋病奈瑟菌pI优势基因型及其G120/A121突变与耐药性关系 |
| Correlation between predominant pI genotypes, G120/A121 mutations and drug resistance |
| Received:September 03, 2009 |
| DOI：10.3760/cma.j.issn.0254-6450.2010.04.019 |
| KeyWord: 淋病奈瑟菌|Porin I基因|双重PCR|耐药性 |
| English Key Word: Neisseria gonorrhoeae|Porin I gene|Double polymerase chain reaction|Drug resistanc |
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| Abstract: |
| 目的 分析浙江省上虞地区淋病奈瑟菌临床菌株外膜孔蛋白优势基因型及其G120和A121突变与耐药性关系。方法 建立能同时检测pIA+和pIB+基因的双重PCR。目的扩增产物T-A克隆后测序, 以分析G120和A121突变情况及证实双重PCR的检测特异性。采用酸性纸片法和二倍琼脂稀释法, 分别检测pIA+和pIB+菌株产β-内酰胺酶情况以及对6种抗生素的耐药性。结果 多重PCR可准确地对所有受检淋病奈瑟菌临床菌株进行porinI(pI)基因分型, 其检测灵敏度为1ng DNA模板。 116株淋病奈瑟菌临床菌株中, 30.2%(35/116)为pIA+菌株, 69.8%(81/136)为pIB+菌株。所有pIA+菌株出现G120D/A121G双突变(88.6%)或A121G单突变(11.4%), 98.8% pIB+菌株出现G120K/A121D(65.0%)、G120K/A121G或G120N/A121D(13.8%)双突变以及G120D/N/K单突变(21.3%)。34.5%(40/116)菌株产β-内酰胺酶, 其中pIA+菌株产酶率(20%)明显低pIB+菌株(40.7%)(P<0.05)。上述临床菌株对青霉素、四环素、环丙沙星和阿奇霉素耐药率高达75.0%～90.5%, 仅有3株菌株对头孢曲松耐药, 未发现大观霉素耐药菌株。100%和71.4%不产β-内酰胺酶、G120和/或A121突变pIA+菌株分别对青霉素和四环素敏感, 但不产β-内酰胺酶G120和/或A121突变的pIB+菌株对该2种抗生素耐药率均为100%。结论 所建立的多重PCR可用于淋病奈瑟菌pI基因快速和准确分型。上虞地区流行的淋病奈瑟菌主要携带pIB基因。大观霉素和头孢曲松仍可作为治疗淋病的首选药物。G120和/或A121突变增强对青霉素和四环素耐药性仅限于β-菌株。 |
| English Abstract: |
| Objective To analyze the predominant genotypes of outer membrane porin I(PI) and to determine the correlation between G120 as well as A121 mutations in PI proteins and drug resistance in Neisseria gonorrhoeae isolates in the local area. Methods A double PCR to simultaneously detect both pIA and pIB genes, was established in this study. The target amplification products were T-A cloned and then sequenced to determine the mutations at G120, A12l and the specificity of double PCR. By using acidity slip method and double agar dilution method, the β-lactamasep roduction and resistance to six antibiotics of pIA+ and pIB+ gonococeal isolates were detected. Results Double PCR could be used to accurately genotyping pI genes in all the tested gonococcal isolates with the sensitivity of lng DNA template. In the 116 N.gonorrhoeae isolates, 30.2% (35/116) were pIA+ strains and 69.8% (81/136) were pIB+ strains. All the pIA+ strains presented G120D/A121G double mutations (88.6%) or A12IG single mutation (11.4%). 98.8% of the pIB+ strainsp resented G120K/A12lD(65.0%), G120K/A121G or G120N/A12lD (13.8%) double mutations, and G120D/N/K single mutation(21.3%). 34.5% (40/116) of the isolates produced B-lactamase, and the enzyme-produced rate (20%) in pIA+ strains was significantly lower than that in pIB+ strains (40.7%) with P<0.05. No spectinomyein-resistant swains were identified but three ceftriaxone-resistant strains were presented. However, the resistance ratios to penicilin, tetramycin, ciprofloxacin and azithromycin of all the isolates were as high as 75.0%-90.5%. 100% and 71.4% of the pIA+ strains without β-lactamase production and with G120 and/or A121 mutations were sensitive to penicillin and terramycin, respectively. On the contrast, 100% of the pIB+ strains without β-lactamase production and with G120 and/or A121 mutations were resistant to both the two antibiotics. Conclusion The established double PCR method could be used for fast and accurate genotyping of N.gonorrhoeae pI genes. The N.gonorrhoeae strains prevalent in the local areas mainly possessed pIB gene. Both spectinomycin and ceftriaxone could still be chosen to trcat gonorrhea. The resistance enhancement caused by G120 and/or A121 mutations to penicillin and tetramycin was only presented in pIB+gonococci. |
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