荧光熔解曲线法和GeneXpert MTB/RIF技术对结核分枝杆菌复合群和利福平耐药检测结果的比较

刘𤧟; 梁颖; 伍仕敏; 章敏

doi:10.13201/j.issn.1004-2806.2022.02.007

荧光熔解曲线法和GeneXpert MTB/RIF技术对结核分枝杆菌复合群和利福平耐药检测结果的比较

- 1.
  武汉市金银潭医院检验科(武汉，430032)
- 2.
  长江航运总医院检验科

详细信息

通讯作者: 伍仕敏，E-mail：mintyrain@126.com

中图分类号: R457.1

收稿日期: 2021-06-22

修回日期: 2021-07-26

刊出日期: 2022-02-01

Results comparison of fluorescence PCR melting curve method and GeneXpert MTB/RIF technology on detecting Mycobacterium tuberculosis complex and rifampicin resistance

- 1.
  Department of Laboratory Medicine, Wuhan Jinyintan Hospital, Wuhan, 430032, China
- 2.
  Department of Laboratory Medicine, Yangtze River Shipping General Hospital

More Information

Corresponding author: WU Shimin, E-mail: mintyrain@126.com

Received Date: 22 June 2021

Revised Date: 26 July 2021

Publish Date: 01 February 2022

摘要

摘要: 目的比较荧光熔解曲线法和GeneXpert MTB/RIF技术对不同类型样本结核分枝杆菌复合群和利福平耐药性的检测结果，旨在为临床不同类型样本如何选择试验方法提供参考，以提高检出率。方法收集2019年8月—2020年7月收治的疑似和确诊结核病患者的临床资料和实验室数据，回顾性分析荧光熔解曲线法和GeneXpert MTB/RIF技术检测结核分枝杆菌复合群和利福平耐药情况。结果荧光熔解曲线法共检测样本937例，检出结核分枝杆菌复合群282例，阳性率30.10%(282/937)；其中呼吸道样本、术后标本阳性率分别为29.07%(252/867)、57.69%(30/52)，其他类型样本未检测出阳性。GeneXpert MTB/RIF共检测样本3807例，检出结核分枝杆菌复合群1315例，阳性率34.54%(1315/3807)；其中呼吸道样本、术后标本、胸水、腹水、脑脊液、其他体液、24 h尿沉渣以及粪阳性率分别为36.05%(1099/2799)、75.09%(202/269)、14.34%(71/495)、9.62%(5/52)、10.87%(15/138)、22.22%(4/18)、29.63%(8/27)和11.11%(1/9)。GeneXpert MTB/RIF技术检测结核分枝杆菌复合群的阳性率明显高于荧光熔解曲线法(χ²=6.655，P=0.010)，其中，呼吸道样本、术后标本的检出率明显高于其他样本类型(χ²=29.584，P< 0.01；χ²=6.584，P=0.017)。133例患者同时进行了荧光熔解曲线法、GeneXpert MTB/RIF技术和结核菌培养及表型药敏试验，结果表明表型药敏检测利福平敏感率为68.42%(91/133)、利福平耐药率为31.58%(42/133)；荧光熔解曲线法检测结果为利福平敏感率为63.91%(85/133)、利福平耐药率为30.83%(41/133)；GeneXpert MTB/RIF技术检测结果为利福平敏感率为66.92%(89/133)、利福平耐药率为31.58%(42/133)；2种分子药敏检测结果与表型药敏检测结果比较，差异无统计学意义(P>0.05)。结论 GeneXpert MTB/RIF技术对于临床不同类型的样本结核分枝杆菌复合群的检出率明显优于荧光熔解曲线法，但2种方法检测利福平耐药得到的结果与表型药敏无差异。特别是GeneXpert MTB/RIF技术，因其操作高度自动化，对结核分枝杆菌复合群的检出率较高，更加适合结核患者病原学快速诊断的需求。
- 荧光熔解曲线法 /
- GeneXpert MTB/RIF技术 /
- 结核分枝杆菌符合群 /
- 利福平 /
- 耐药
Abstract: Objective To provide references for options of clinical detection methods in different samples by comparing the results of fluorescence PCR melting curve and GeneXpert MTB/RIF on detecting Mycobacterium tuberculosis complex(MTC) and rifampicin resistance, so as to improve the detection rate of tuberculosis.Methods Clinical and laboratory data of suspected and confirmed tuberculosis patients admitted to our hospital from August 2019 to July 2020 were collectde, and the detection results of MTC and rifampicin resistance which detected by fluorescence melting curve and GeneXpert MTB/RIF were retrospectively analyzed.Results A total of 937 samples were detected by the fluorescence melting curve, and 282 cases of MTC were detected with a positive rate of 30.10%(282/937), of which the positive rates of samples in the respiratory tract and postoperative samples were 29.07%(252/867) and 57.69%(30/52), respectively. There was no positive result in other samples. A total of 3807 samples were detected by GeneXpert MTB/RIF, and 1315 cases of MTC were detected with a positive rate of 34.54%(1315/3807). The positive rates of samples in the respiratory tract, postoperative specimens, pleural fluid, ascites, cerebrospinal fluid, other body fluids, 24-hour urine sediment, and feces were 36.05%(1099/2799), 75.09%(202/269), 14.34%(71/495), 9.62%(5/52), 10.87%(15/138), 22.22%(4/18), 29.63%(8/27) and 11.11%(1/9), respectively. The positive rate of GeneXpert MTB/RIF in detecting MTC was significantly higher than that of fluorescent PCR melting curve(χ²=6.655,P=0.010). Among them, the detection rate of samples in the respiratory tract and postoperative specimens was significantly higher than that of other samples(χ²=29.584,P< 0.01;χ²=6.584,P=0.017). A total of 133 patients were simultaneously detected by fluorescence melting curve, GeneXpert MTB/RIF and tuberculosis culture and phenotypic drug sensitivity test. The results showed that the rate of rifampin sensitivity was 68.42%(91/133) and the rate of rifampicin resistance was 31.58%(42/133) in the phenotypic drug sensitivity test, and the rate of rifampicin sensitivity was 63.91%(85/133) and the rate of rifampicin resistance was 30.83%(41/133) in fluorescence melting curve. And the results showed that the rate of rifampicin sensitivity was 66.92%(89/133) and the rate of rifampicin resistance 31.58%(42/133) in GeneXpert MTB/RIF. The results between the two molecular drug sensitivity tests and the phenotypic drug sensitivity test had no statistical difference(P>0.05).Conclusion The detection rate of GeneXpert MTB/RIF for different types of clinical samples of MTC is significantly higher than the fluorescence melting curve, and there is no difference between the results obtained by the two molecular methods for detecting rifampicin resistance and the phenotypic drug sensitivity. Particularly, because of the highly automated operation of the GeneXpert MTB/RIF and its higher detection rate of MTC, it may be more suitable for the need of rapid pathogenic diagnosis in tuberculosis patients.
- fluorescence melting curve /
- GeneXpert MTB/RIF /
- mycobacterium tuberculosis complex /
- rifampicin /
- drug resistance

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表 1 2种检测方法检出结核分枝杆菌复合群的阳性率

样本类型	荧光熔解曲线法			GeneXpert MTB/RIF技术
样本类型	检测数	阳性数	阳性率/%	检测数	阳性数	阳性率/%
呼吸道样本	867	252	29.07	2799	1099	36.05
术后标本	52	30	57.69	269	202	75.09
胸水	10	0	/	495	71	14.34
腹水	4	0	/	52	5	9.62
脑脊液	3	0	/	138	15	10.87
其他体液	/	/	/	18	4	22.22
尿	1	0	/	27	8	29.63
粪	/	/	/	9	1	11.11
合计	937	282	30.1	3807	1315	34.54
吸道样本包括痰及肺泡灌洗液；术后标本(包括肺组织、淋巴结、脓液、分泌物等)；其他体液包括关节腔液、胸腔积液、腹腔积液、心包积液、引流液。

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