基质辅助激光解吸电离飞行时间质谱鉴定非结核分枝杆菌及药敏试验分析

周亚娣; 金法祥

doi:10.16766/j.cnki.issn.1674-4152.002646

基质辅助激光解吸电离飞行时间质谱鉴定非结核分枝杆菌及药敏试验分析

doi: 10.16766/j.cnki.issn.1674-4152.002646

周亚娣^,,
金法祥

绍兴文理学院附属医院(绍兴市立医院)检验科，浙江绍兴 312000

基金项目:

浙江省医药卫生科技计划项目 2020KY334

详细信息

通讯作者:
周亚娣, E-mail: 1071716897@qq.com

中图分类号: R378.91 R446
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- 文章访问数: 147
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- 被引次数: 0
出版历程
- 收稿日期: 2022-03-04
- 网络出版日期: 2022-11-29

Identification of nontuberculous mycobacteria by matrix-assisted laser desorption ionization time-of-flight mass spectrometry and analysis of drug-sensitivity test

ZHOU Ya-di^,,
JIN Fa-xiang

Department of Clinical Laboratory, Affiliated Hospital of Shaoxing University (Shaoxing Municipal Hospital), Shaoxing, Zhejiang 312000, China

摘要

摘要: 目的研究非结核分枝杆菌(NTM)基质辅助激光解析电离飞行时间质谱(MALDI-TOF MS)分型结果，比较不同NTM菌种耐药情况，为临床合理用药提供依据。方法回顾性分析2019年3月—2021年6月绍兴市区结核病定点医院(绍兴文理学院附属医院)NTM菌株相关资料，将来源为相同患者的菌株资料排除后，剩余128株NTM菌株，用MALDI-TOF MS技术对NTM菌种进行分型，同时进行基因测序分型，以基因测序分型结果为金标准，计算出MALDI-TOF MS技术分型结果的准确率，并对分型后排名前三的NTM进行药敏试验分析。结果 MALDI-TOF MS技术对NTM分型结果的准确率为98.4%。胞内分枝杆菌对阿米卡星、克拉霉素、利奈唑胺、莫西沙星和利福平的敏感率分别为98.5%、89.4%、86.4%、83.3%和62.1%；鸟分枝杆菌对阿米卡星、克拉霉素、利奈唑胺、莫西沙星和利福平的敏感率分别为100.0%、92.9%、100.0%、82.1%和89.3%；两者对米诺环素的耐药率分别为90.9%、82.1%。堪萨斯分枝杆菌对阿米卡星、克拉霉素、利福平的敏感率分别为100.0%、100.0%、95.2%。结论 MALDI-TOF MS技术对NTM分型的准确率高，胞内分枝杆菌、鸟分枝杆菌、堪萨斯分枝杆菌，这3种NTM对阿米卡星和克拉霉素的敏感率高，阿米卡星和克拉霉素可作为临床治疗NTM的首选药物。
- 基质辅助激光解析电离飞行时间质谱技术 /
- 非结核分枝杆菌 /
- 快速分型 /
- 药敏试验
Abstract: Objective To study the typing results of matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) of nontuberculous mycobacteria (NTM), compare the drug resistance of different NTM strains, and provide a basis for clinical rational drug use. Methods The data on NTM strains in Shaoxing City Tuberculosis Designated Hospital (Affiliated Hospital of Shaoxing College of Arts and Sciences) from March 2019 to June 2021 were retrospectively analyzed. After excluding the strain data from the same patient, the remaining 128 NTM strains were classified by MALDI-TOF MS technology. Genotyping was performed at the same time, and the results of genotyping were used as the gold standard. The accuracy of MALDI-TOF MS typing results was calculated, and drug susceptibility tests were performed on the top three NTMs after typing. Results The accuracy of NTM typing by MALDI-TOF MS was 98.4%. The sensitivity rates of mycobacterium intracellular to amikacin (AK), clarithromycin (CLA), linezolid (LZD), moxifloxacin (MXF) and rifampicin (RFP) were 98.5%, 89.4%, 86.4%, 83.3% and 62.1%, respectively. The sensitivity rates of mycobacterium avium to AK, CLA, LZD, MXF and RFP were 100.0%, 92.9%, 100.0%, 82.1% and 89.3%, respectively. The drug resistance rates of the above strains to minocycline (MH) were 90.9% and 82.1%, respectively. The sensitivity rates of mycobacterium Kansas to AK, CLA and RFP were 100.0%, 100.0% and 95.2%, respectively. Conclusion MALDI-TOF MS has high accuracy in NTM typing, and mycobacterium intracellular, Mycobacterium avium and Mycobacterium Kansas are highly sensitive to AK and CLA. AK and CLA can be used as the first choice for clinical treatment of NTM.
- Matrix-assisted laser desorption ionization time-of-flight mass spectrometry /
- Nontuberculosis mycobacteria /
- Rapid typing /
- Drug-sensitivity test

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表 1 66株胞内分枝杆菌药敏试验结果

Table 1. Results of drug susceptibility test of 66 strains of intracellular Mycobacterium

药物	MIC₅₀ (μg/mL)	MIC₉₀ (μg/mL)	敏感[株(%)]	中度敏感[株(%)]	耐药[株(%)]
阿米卡星	8.000	16.000	65(98.5)	-	1(1.5)
克拉霉素	0.500	8.000	59(89.4)	2(3.0)	5(7.6)
利奈唑胺	0.500	8.000	57(86.4)	-	9(13.6)
莫西沙星	0.125	＞16.000	55(83.3)	4(6.1)	7(10.6)
利福平	≤1.000	16.000	41(62.1)	-	25(37.9)
米诺环素	＞128.000	＞128.000	5(7.6)	1(1.5)	60(90.9)
注：“-”表示该药物在CLSI标准上没有药物临界折点; MIC₅₀为抑制50%细菌生长最低抑菌浓度; MIC₉₀为抑制90%细菌生长最低抑菌浓度。

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表 2 28株鸟分枝杆菌药敏试验结果

Table 2. Drug susceptibility test results of 28 strains of Mycobacterium avium

药物	MIC₅₀ (μg/mL)	MIC₉₀ (μg/mL)	敏感[株(%)]	中度敏感[株(%)]	耐药[株(%)]
阿米卡星	8.000	16.000	28(100.0)	-	-
克拉霉素	0.500	8.000	26(92.9)	2(7.1)	-
利奈唑胺	0.500	8.000	28(100.0)	-	-
莫西沙星	0.125	＞16.000	23(82.1)	2(7.1)	3(10.7)
利福平	≤1.000	16.000	25(89.3)	-	3(10.7)
米诺环素	＞128.000	＞128.000	1(3.6)	4(14.3)	23(82.1)
注：“-”表示该药物在CLSI标准上没有药物临界折点; MIC₅₀为抑制50%细菌生长最低抑菌浓度; MIC₉₀为抑制90%细菌生长最低抑菌浓度。

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表 3 21株堪萨斯分枝杆菌药敏试验结果

Table 3. Drug susceptibility test results of 21 strains of Mycobacterium Kansas

药物	MIC₅₀ (μg/mL)	MIC₉₀ (μg/mL)	敏感[株(%)]	中度敏感[株(%)]	耐药[株(%)]
阿米卡星	≤0.500 0	8.000 0	21(100.0)	-	-
克拉霉素	≤0.062 5	1.000 0	21(100.0)	-	-
利福平	≤1.000 0	≤1.000 0	20(95.2)	-	1(4.8)
注：“-”表示该药物在CLSI标准上没有药物临界折点。MIC₅₀为抑制50%细菌生长最低抑菌浓度; MIC₉₀为抑制90%细菌生长最低抑菌浓度。

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