scholarly journals Novel Mutations Detected from Drug Resistant Mycobacterium Tuberculosis Isolated from North East of Thailand

Author(s):  
Ei Phoo Thwe ◽  
Wises Namwat ◽  
Porntip Pinlaor ◽  
Kulrattana Rueangsak ◽  
Arunnee Sangka

Abstract The emergence of drug-resistant tuberculosis is a major global public health threat. Thailand is one of the top 14 countries with high tuberculosis, tuberculosis co-infected with Human Immunodeficiency Virus (HIV) and multi-drug resistant tuberculosis rates. Immediate detection of drug-resistant tuberculosis is necessary to reduce mortality and morbidity by effectively providing treatment to ameliorate the formation of resistant strains. Limited data exist of mutation profiles in Northeastern Thailand. Here, 65 drug-resistant Mycobacterium tuberculosis isolates were used to detect mutations by polymerase chain reaction (PCR) and DNA sequencing. In the katG gene, mutations occurred in 47(79.7%) among 59 isoniazid resistant samples. For rpoB gene, 31 (96.9%) were observed as mutations in 32 rifampicin resistant isolates. Of 47 katG mutation samples, 45 (95.7%) had mutations in katG 315 codon and 2 (4.3%) showed novel mutations at katG 365 with amino acid substitution of CCG-CGG (Pro-Arg). Moreover, 18 (58.1%) mutations at rpoB 531, 11 (35.5%) mutations at rpoB 516, 10 (32.3%) mutations at rpoB 526 and 1 (3.2%) mutation at rpoB 533 were found between 31 rifampicin resistant samples. Common and novel mutations of the rpoB and katG genes in Northeastern Thailand were generated. DNA sequencing showed high accuracy, while conventional polymerase chain reaction could be applied as an initial marker for screening rifampicin and isoniazid drug-resistant Mycobacterium tuberculosis in Northeastern Thailand. Mutations reported here should be considered when developing new molecular diagnostic methods for implementation in Northeastern Thailand.

Author(s):  
Matthew J. Binnicker ◽  
Glenn D. Roberts ◽  
Nancy L. Wengenack

This chapter reviews diagnostic methods and tests for identifying mycobacterial and fungal organisms. Diagnostic methods include direct examination, staining, culture, molecular identification, DNA sequencing, chromatography, polymerase chain reaction, and immunodiagnostics. Organisms reviewed include Mycobacterium tuberculosis, M kansasii, M marinum, M leprae, M avium, and other mycobacteria; Aspergillus spp; Histoplasma spp; Coccidioides spp; Blastomyces spp; Candida spp; Fusarium spp; Trichophyton spp; and other fungi.


2012 ◽  
Vol 11 (4) ◽  
pp. 322-330 ◽  
Author(s):  
Shamima Islam ◽  
Farjana Rahman ◽  
Saurab Kisore Munshi ◽  
Jewel Ahmed ◽  
S M Mostafa Kamal ◽  
...  

Objective: Drug resistant tuberculosis has long been a common problem prevailing in developing countries including Bangladesh. Present study focused on the rapid identification of live Mycobacterium tuberculosis among treatment failure cases.Materials and Methods: Sputum samples from a total of 100 category-I and category-II treatment failure cases, assumed as multidrug resistant tuberculosis, were studied through fluorescein diacetate (FDA) staining under light emitting diode (LED) fluorescence microscope. Considering culture method as gold standard, we also compared the results of FDA staining with that of auramine O staining.Results: A total of 85% acid-fast bacilli were detected by FDA staining, 82% by auramine O staining and a total of 85% isolates were detected in Lowenstein-Jensen (LJ) culture. The sensitivity of FDA staining (96.47%) was estimated to be slightly higher than that of auramine O staining (91.76%). Moreover,76.47% cases were detected as multidrug resistant tuberculosis (MDR-TB). Conclusion: Taken together, FDA staining method has been proposed to be appropriate for the rapid diagnosis of drug resistant tuberculosis. DOI: http://dx.doi.org/10.3329/bjms.v11i4.12605 Bangladesh Journal of Medical Science Vol. 11 No. 04 Oct’12


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