荧光PCR熔解曲线法分析宜昌地区非结核分枝杆菌感染的流行特征
作者:
作者单位:

1.宜昌市第三人民医院检验科;2.武汉大学公共卫生学院

作者简介:

通讯作者:

江琦  E-mail: jiang.qi@whu.edu.cn

中图分类号:

+2  R378.91]]>

基金项目:

湖北省宜昌市医疗卫生研究项目(A22-2-076)


Epidemic characteristics of non-tuberculous mycobacterial infection in Yichang area by fluorescence polymerase chain reaction melting curve method
Author:
Affiliation:

1.Department of Laboratory Medicine, The Third People's Hospital of Yichang, Yichang 443000, China;2.School of Public Health, Wuhan University, Wuhan 430061, China

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    摘要:

    目的 应用荧光PCR熔解曲线法分析宜昌地区非结核分枝杆菌(NTM)的感染趋势、分布特征、流行状况,为该地区NTM感染的防治提供参考依据。 方法 收集2016—2020年间宜昌地区在某院就诊的疑似结核病患者的胸腔积液、腹腔积液、痰、支气管肺泡灌洗液标本,经萋-尼抗酸染色涂片、MGIT 960快速培养和改良罗氏培养检出的所有分枝杆菌阳性标本进一步经荧光PCR熔解曲线法鉴定结核分枝杆菌(MTB)及NTM菌种。以全基因组测序(WGS)为金标准,对比分析荧光PCR熔解曲线法鉴定MTB及NTM的准确性,总结该地区NTM感染患者的临床特征。 结果 共收集7 496份分枝杆菌阳性标本,经荧光PCR熔解曲线法鉴定,检出6 997株(93.34%)MTB与499株(6.66%)NTM。NTM菌种鉴定共确定469株(93.99%)NTM菌种,其余30株(6.01%)为未知菌种。检出最多的NTM为胞内分枝杆菌(189, 37.88%),其次依次为脓肿分枝杆菌(68株, 13.63%)、戈登分枝杆菌(25株, 5.01%)。选择荧光PCR熔解曲线法鉴定的22株MTB和18株NTM行WGS,符合率为98.15%,灵敏度为99.80%,特异度为93.54%。分析NTM流行特征发现,该地区NTM检出率逐年升高,自2016年的3.56%上升至2020年的13.21%。NTM阳性患者中,男性占57.52%(287/499),52.30%的患者(261/499)集中在55~74岁年龄段。 结论 宜昌地区NTM流行率呈快速增加趋势,以脓肿、戈登、堪萨斯分枝杆菌为主,荧光PCR熔解曲线法可有效快速辅助诊断。老年人NTM感染风险高,临床上鉴别诊断NTM意义重大。

    Abstract:

    Objective To analyze the infection trend, distribution characteristics and epidemic status of non-tuberculous mycobacterial (NTM) infection in Yichang area by fluorescence polymerase chain reaction (PCR) melting curve method, providing reference for the prevention and treatment of NTM infection in this area. Methods Pleural effusion, peritoneal effusion, sputum and bronchoalveolar lavage fluid from patients with suspected tuberculosis in a hospital in Yichang area from 2016 to 2020 were collected. All Mycobacterium positive specimens detected by Ziehl-Neelsen acid-fast staining smear, MGIT 960 rapid culture and modified Loewenstein-Jenson medium culture were further identified Mycobacterium tuberculosis (MTB) and NTM by fluorescence PCR melting curve method. With whole genome sequencing (WGS) as the gold standard, the accuracy of fluorescence PCR melting curve method in identifying MTB and NTM was compared and analyzed, and clinical characteristics of NTM infected patients in Yichang area were summarized. Results A total of 7 496 Mycobacterium positive specimens were collected. After identified by fluorescence PCR melting curve method, 6 997 (93.34%) MTB strains and 499 (6.66%) NTM strains were detected. A total of 469 (93.99%) NTM strains were identified, while the remaining 30 (6.01%) strains were unknown strains. The most commonly detected NTM was Mycobacterium intracellulare (n=189, 37.88%), followed by Mycobacterium abscessus (n=68, 13.63%) and Mycobacterium gordonae (n=25, 5.01%). 22 MTB strains and 18 NTM strains identified by fluorescence PCR melting curve method were selected to perform WGS, and the coincidence rate, sensitivity, and specificity were 98.15%, 99.80%, and 93.54%, respectively. The epidemic characteristics of NTM showed that the detection rate of NTM in this area has been increasing year by year, from 3.56% in 2016 to 13.21% in 2020. Among NTM positive patients, 57.52% (287/499) were male, 52.30% (261/499) of the patients were aged 55-74 years old. Conclusion The prevalence of NTM in Yichang area presents a rapid increasing trend, with Mycobacterium abscessus, Mycobacterium gordonae and Mycobacterium kansasii as the main pathogens. Fluorescence PCR melting curve method can effectively and quickly assist in diagnosis. The risk of NTM infection in elderly people is high, and differential diagnosis is of great significance in clinical practice.

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胡晓红,韩宇,庄倩,等.荧光PCR熔解曲线法分析宜昌地区非结核分枝杆菌感染的流行特征[J]. 中国感染控制杂志,2023,(8):939-944. DOI:10.12138/j. issn.1671-9638.20234121.
Xiao-hong HU, Yu HAN, Qian ZHUANG, et al. Epidemic characteristics of non-tuberculous mycobacterial infection in Yichang area by fluorescence polymerase chain reaction melting curve method[J]. Chin J Infect Control, 2023,(8):939-944. DOI:10.12138/j. issn.1671-9638.20234121.

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  • 收稿日期:2023-02-17
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  • 在线发布日期: 2024-04-28
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