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| 云南昭通地区肺结核患者中结核分枝杆菌基因多样性与耐药分子特征的相关性研究 |
| Study on the correlation between MTB genetic diversity and drug resistance molecular characteristics in pulmonary tuberculosis patients in Zhaotong, Yunnan Province |
| 投稿时间：2024-02-19 修订日期：2024-04-25 |
| DOI： |
| 中文关键词: 云南昭通地区 结核分枝杆菌 基因多样性 MIRU-VNTR 24位点分型法 Mcspoligotping |
| 英文关键词: Zhaotong, Yunnan Province Mycobacterium tuberculosis genetic diversity MIRU-VNTR 24-site typing Mcspoligotping |
| 基金项目:云南省科技厅科技计划项目（202201AY070001-219） |
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| 摘要点击次数: 30 |
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| 中文摘要: |
| 目的 旨在调查云南昭通地区结核分枝杆菌（mycobacterium tuberculosis, MTB）临床菌株的遗传多样性和耐药分子特征。方法 2022年9月至2023年8月，从云南省昭通市第一人民医院涂阳PTB患者中共采集了298份MTB菌株样本。使用MeltPro TB测定法通过荧光PCR熔解曲线法鉴定所有分离株。比例法用于测定结核分枝杆菌的体外药物敏感性。使用多色熔解曲线分析技术进行Spoligotping分析。通过分枝杆菌穿插重复单位可变数串联重复（MIRU-VNTR）24位点分型法进行基因分型。使用PCR和靶向测序检测耐药基因突变。结果 北京菌株（69.80%，208/298）是云南MTB菌株中最主要的谱系。其他谱系包括T（17/298=5.70%）、LAM（10/298=3.36%）、MANU2（2/298=0.67%）、CAS（1/298=0.34%）、NEW-1（25/298=8.39%）和SITVITWEB数据库中未发现的孤儿型（35/298=11.74%）。北京菌株对左氧氟沙星的耐药性略高于非北京菌株（P=0.042），而2种基因型菌株中异烟肼、利福平、乙胺丁醇、链霉素、吡嗪酰胺、卡那霉素等其他耐药谱特征无统计学差异（P＞0.05）。MIRU-VNTR 24位点分型分析结果表明，33个菌株可分为13株聚类，其聚类率为39.39%。根据Hunter-Gaston判别指数，每个等位基因的多样性在0.00至0.814之间存在显著差异，其中18个位点对非北京菌株具有中高鉴别能力。但只有10个位点对北京菌株具有中高鉴别能力。结论 MTB菌株在中国云南昭通地区表现出较高的遗传多样性，北京菌株是MTB和耐药结核病菌株的优势谱系。 |
| 英文摘要: |
| Objective To investigate the genetic diversity and drug resistance of clinical strains of Mycobacterium tuberculosis (MTB) in Zhaotong, Yunnan Province. Methods From September 2022 to August 2023, 298 clinical isolates of MTB from smear-positive pulmonary tuberculosis patients in the First People"s Hospital of Zhaotong were collected. MeltPro TB assay was used to identify all isolates by fluorescence PCR meltdown curve. Proportional method was used to determine the drug sensitivity of mycobacterium tuberculosis in vitro. Spoligotping analysis was performed using polychromatic melting curve analysis technique. Genotyping was performed by Mcspoligotping and Mycobacteria interspersing repetitive unit Variable Tandem repeats (MIRU-VNTR) 24-site typing. Drug-resistant gene mutations were detected using PCR and targeted sequencing. Results Beijing strain (69.80%, 208/298) was the most dominant lineage of MTB strains in Yunnan. Other lineages included T (17/298=5.70%), LAM (10/298=3.36%), MANU2 (2/298=0.67%), CAS (1/298=0.34%), NEW-1 (25/298=8.39%), and orphan type not found in SITVITWEB database (35/ 298=11.74%). The resistance of Beijing strain to levofloxacin was slightly higher than that of non-Beijing strain (P=0.042), but there was no significant difference in isoniazid, rifampicin, ethambutol, streptomycin, pyrazinamide, kanamycin and other resistance profiles between the two genotypes (P＞0.05). The results of MIRU-VNTR 24 locus typing showed that the 33 strains could be grouped into 13 strains, and the clustering rate was 39.39%. According to the Hunter-Gaston discriminant index, the diversity of each allele varied significantly from 0.00 to 0.814, with 18 loci having medium-high discriminability against non-Beijing strains. However, only 10 loci had medium-high discriminability against Beijing strain. Conclusion MTB strains in Zhaotong, Yunnan, China, show a high genetic diversity. Beijing strains are the dominant lineage of MTB and drug-resistant TB strains |
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