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Detecting drug resistant genetic mutation among pneumoconiosis patients complicated with tuberculosis in Mycobacterium tuberculosis L-forms application of PCR-SSCP technique in Huainan mining district
作者姓名:Jun Lu  Shan Jiang  Song Ye  Chaopin Li
作者单位:[1]Medical College, Anhui University of Science & Technology, Huainan 232001, China [2]Department of Mining Engineering, Huainan Vocational & Technical College, Huainan 232001, China [3]Wannan Medical College, Wuhu 241001, China
基金项目:This work was supported by the Youth Natural Science Foundation of Anhui University of Science & Technology (200537)
摘    要:Objective: To study the relationship between drug resistant genetic mutation and drug resistance in Mycobacterium tuberculosis L-form, discuss the internal relationship between drug resistances and drug-resistant related genes and explore the value of PCR- SSCP to clinical application. Methods: A total of 52 clinically isolated strains of tuberculosis L-form were collected among 97 pneumoconiosis patients complicated with tuberculosis. The gene mutations of katG, rpoB and rpsL were detected by PCR-SSCP, and the results were compared with those analyzed by traditional antimicrobial susceptibility test(AST). Results: The gene muta- tion rates of katG, rpoB and rpsL by PCR-SSCP were respectively 57.70% (30/52), 65.38% (32/52) and 40.38% (21/52). The rate of reversion was 78.85%(41/52) and the result of drag-resistant genes was invariable. The results of AST showed that there were 40 (76.92%) multi-drug resistant strains in 52 clinically isolated strains. The number for three-drug resistant strain was 21 (40.38%) and that of two-drug resistant was 19(36.54%), but only 12(23.08%) strains were one drug resistant. The rate of total drug-resistance was 100%, but there were 15 strains of allied mutation of three genes, 16 of two mutations and 6 of only one by PCR-SSCP. The coincidences were respectively 71.43%, 84.12% and 50.00%. Then there was no significant difference between the allied mutations of multi-drug resistant gene and the mutations of only one drug resistant gene (P 〉 0.05). Conclusion: PCR-SSCP technique has a higher sensibility and specificity to detect the genes of katG, rpoB and rpsL in tuberculosis L-form among pneumoconiosis complicated with tuberculosis,and the detecting rate of two drug resistant strains and three drug resistant strains was higher. The combined application of PCR-SSCP and AST has advantages at earlier diagnosis and guidance of clinical medications.

关 键 词:淮南矿区  肺尘症患者  并发  肺结核  结核分支杆菌  耐药基因突变  检测  PCR-SSCP
收稿时间:2007-01-23

Detecting drug resistant genetic mutation among pneumoconiosis patients complicated with tuberculosis in Mycobacterium tuberculosis L-forms application of PCR-SSCP technique in Huainan mining district
Jun Lu,Shan Jiang,Song Ye,Chaopin Li.Detecting drug resistant genetic mutation among pneumoconiosis patients complicated with tuberculosis in Mycobacterium tuberculosis L-forms application of PCR-SSCP technique in Huainan mining district[J].Acta Universitatis Medicinalis Nanjing,2007,21(3):180-184.
Authors:Jun Lu  Shan Jiang  Song Ye and Chaopin Li
Institution:Medical College, Anhui University of Science & Technology, Huainan 232001, China;Department of Mining Engineering, Huainan Vocational & Technical College, Huainan 232001, China;Medical College, Anhui University of Science & Technology, Huainan 232001, China;Wannan Medical College, Wuhu 241001, China
Abstract:Objective: To study the relationship between drug resistant genetic mutation and drug resistance in Mycobacterium tuberculosis L-form, discuss the internal relationship between drug resistances and drug-resistant related genes and explore the value of PCRSSCP to clinical application. Methods: A total of 52 clinically isolated strains of tuberculosis L-form were collected among 97pneumoconiosis patients complicated with tuberculosis. The gene mutations of katG, rpoB and rpsL were detected by PCR-SSCP,and the results were compared with those analyzed by traditional antimicrobial susceptibility test(AST). Results: The gene mutation rates of katG, rpoB and rpsL by PCR-SSCP were respectively 57.70% (30/52), 65.38% (32/52) and 40.38% (21/52). The rate of reversion was 78.85%(41/52) and the result of drug-resistant genes was invariable. The results of AST showed that there were 40 (76.92%) multi-drug resistant strains in 52 clinically isolated strains. The number for three-drug resistant strain was 21(40.38%) and that of two-drug resistant was 19(36.54%), but only 12 (23.08%) strains were one drug resistant. The rate of total drug-resistance was 100%, but there were 15 strains of allied mutation of three genes, 16 of two mutations and 6 of only one by PCR-SSCP. The coincidences were respectively 71.43%, 84.12% and 50.00%. Then there was no significant difference between the allied mutations of multi-drug resistant gene and the mutations of only one drug resistant gene (P > 0.05). Conclusion: PCRSSCP technique has a higher sensibility and specificity to detect the genes of katG, rpoB and rpsL in tuberculosis L-form among pneumoconiosis complicated with tuberculosis,and the detecting rate of two drug resistant strains and three drug resistant strains was higher. The combined application of PCR-SSCP and AST has advantages at earlier diagnosis and guidance of clinical medications.
Keywords:tuberculosis  Mycobacterium tuberculosis L-form  drug-resistance  katG  rpoB  rpsL  polymerase chain reaction and single-strand conformation polymorphism (PCR-SSCP)  district  mining  Huainan  technique  application  Mycobacterium tuberculosis  complicated  patients  pneumoconiosis  genetic mutation  resistant gene  drug  combined  diagnosis  guidance  clinical  detecting  sensibility  specificity  significant
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