Diagnostic value of melting curve method in detecting resistance of Mycobacterium tuberculosis to rifampicin and isoniazid

doi:10.3969/j.issn.2096-8493.2020.03.008

Abstract

Abstract:

Objective To evaluate the efficacy of fluorescence PCR probe melting curve technique (hereinafter referred to as melting curve technique) in detecting for the resistance of Mycobacterium tuberculosis to rifampicin and isoniazid. Methods Sputum specimens of 832 smear-positive pulmonary tuberculosis patients were cultured in liquid, and the results showed that specimens of 35 cases were contaminated, 52 were culture negative, and 745 were positive. Seven hundred and forty-five positive cultures were then tested with gene chip which found 7 cases of mixed infection, 60 cases of non-mycobacterium tuberculosis, and 678 cases of Mycobacterium tuberculosis complex. The BACTEC MGIT 960 (“MGIT 960”) liquid drug sensitivity test and melting curve method were then performed on specimens of those 678 cases to detect drug resistance of rifampicin and isoniazid. The results of the melting curve method in rifampicin detection were invalid (impossible to tell whether it’s resistant) in 31 cases and isoniazid in 34 cases,for the remaining 639 cases valid results were reported for both rifampicin and isoniazid. Taking MGIT 960 liquid drug sensitivity test method as the golden standard to analyze the performance of melting curve method. Results The rate of concordance, sensitivity, specificity, positive predictive value and negative predictive value to rifampicin detection were 96.2% (615/639), 93.3% (126/135), 97.0% (489/504), 89.4% (126/141) and 98.2% (489/498) respectively. The Kappa value was 0.89. The rate of concordance, sensitivity, specificity, positive predictive value and negative predictive value to isoniazid detection were 93.6% (598/639), 83.1% (157/189), 98.0% (441/450), 94.6% (157/166) and 93.2% (441/473) respectively. The Kappa value was 0.84. Conclusion The melting curve method has high sensitivity and good specificity for detecting rifampicin and isoniazid resistance, thus is of great value for early diagnosis of drug-resistant tuberculosis.

Key words: Tuberculosis,multidrug-resistant, Polymerase chain reaction, Microbial susceptibility tests, DNA probes, Microarray analysis, Comparative study, Data interpretation,statistical

SU Bi-yi, ZHOU De-wang, MA Pin-yun, GUAN Ping, TAN Yao-ju. Diagnostic value of melting curve method in detecting resistance of Mycobacterium tuberculosis to rifampicin and isoniazid[J]. Journal of Tuberculosis and Lung Disease , 2020, 1(3): 245-248. doi: 10.3969/j.issn.2096-8493.2020.03.008

Figures/Tables 2

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药品名称	熔解曲线法	MGIT 960液体药敏试验		敏感度 (%)	特异度 (%)	阳性预测值 (%)	阴性预测值 (%)	Kappa值
药品名称	熔解曲线法	耐药(例)	敏感(例)	敏感度 (%)	特异度 (%)	阳性预测值 (%)	阴性预测值 (%)	Kappa值
利福平	耐药(例)	126	15	93.3	97.0	89.4	98.2	0.89
	敏感(例)	9	489
异烟肼	耐药(例)	157	9	83.1	98.0	94.6	93.2	0.84
	敏感(例)	32	441

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