Application value of multicolor melting curve analysis in detection of pre-extensive drug-resistant pulmonary tuberculosis

doi:10.19982/j.issn.1000-6621.20220418

Abstract

Abstract:

Objective: To evaluate the application value of multicolor melting curve analysis (MMCA) in detection of pre-extensive drug-resistant pulmonary tuberculosis (pre-XDR-PTB). Methods: A total of 129 sputum samples from 129 patients with positive culture and initially identified as multidrug-/rifampicin-resistant pulmonary tuberculosis (MDR/RR-PTB) were collected. All the patients were registered in the Shenzhen Center for Chronic Disease Control and district-level tuberculosis prevention and control institutions from January 2019 to December 2020. The samples were detected by drug susceptibility tests (DST) of three fluoroquinolones (ofloxacin (Ofx), levofloxacin (Lfx), and moxifloxacin (Mfx)) and MMCA. Sanger sequencing was performed as the third-party verification of the results. Using DST as the gold standard, the efficacy of MMCA in detecting fluoroquinolone resistance in patients with MDR/RR-PTB was analyzed. Results: Using DST as the gold standard, the sensitivity, specificity, and Kappa values of the MMCA method for detecting fluoroquinolone resistance in MDR/RR-PTB patients’ samples were 87.0% (40/46), 94.0% (78/83), and 0.813, respectively, for Ofx; 97.3% (36/37), 90.2% (83/92), and 0.822, respectively, for Lfx; 92.3% (24/26), 79.6% (82/103), and 0.565, respectively, for Mfx. If combining the DST results of three fluoroquinolones and defining fluoroquinolone resistance as resistance to any one of the three fluoroquinolones, the sensitivity, specificity, and Kappa value of the MMCA method for detecting fluoroquinolone resistance in MDR/RR-PTB patients’ samples were 85.1% (40/47), 93.9% (77/82), and 0.797, respectively. Sanger sequencing was performed on 52 suspected of drug resistance samples. Among these samples, 96.2% (50/52) had the same sequencing results as MMCA, and 3.8% (2/52) of the samples had mutations in the non-detection region of MMCA. Conclusion: The results of MMCA were consistent with those of DST, and thus MMCA could be used to detect pre-XDR-PTB.

Key words: Mycobacterium tuberculosis, Drug resistance, multiple, bacterial, Microbial sensitivity tests, Molecular diagnostic techniques, Diagnosis, differential

CLC Number:

R52
R825.2

Zhu Yumei, Xia Zihan, Li Jinli, He Huishan, Wang Feng. Application value of multicolor melting curve analysis in detection of pre-extensive drug-resistant pulmonary tuberculosis[J]. Chinese Journal of Antituberculosis, 2023, 45(4): 406-411. doi: 10.19982/j.issn.1000-6621.20220418

Figures/Tables 2

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MMCA检测	表型药敏试验				Sanger测序	标本份数
MMCA检测	FQs	Ofx	Lfx	Mfx	Sanger测序	标本份数
突变型	R	R	R	R	gyrA D94G	12
	R	R	R	R	gyrA D94N	4
	R	R	R	R	gyrA D94H	2
	R	R	R	R	gyrA S91P	2
	R	R	R	R	gyrA G88C	2
	R	R	R	R	gyrA D94Y	1
	R	R	R	R	gyrA A90V	1
	R	R	R	S	gyrA A90V	6
	R	R	R	S	gyrA D94A	2
	R	R	R	S	gyrA D94G	2
	R	R	R	S	gyrA D94N	1
	R	R	R	S	gyrA D94H	1
	R	R	S	S	gyrA D94A	3
	R	R	S	S	gyrA A90V	1
	S	S	S	S	gyrA D94A	3
	S	S	S	S	gyrA A90V	2
野生型	R	R	R	R	gyrB E540D	1
	R	R	S	S	gyrB野生型	5
	R	S	S	R	gyrB E540D	1
	S	S	S	S	未测序(表型药敏试验结果与MMCA检测结果一致)	77