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The Use of the pcr-based dot-blot Hybridization Assay to Detect Resistance Markers to Rifampicin and Streptomycin in Mycobacterium tuberculosis Isolates from the SW Region of Cameroon

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Journal of Tuberculosis Research 2016年第2期4卷 72-79页

作者： Irene Ane-Anyangwe Wilfred Fon Mbacham Henry Dilonga Meriki Teyim Pride Theresa Nkuo-Akenji Veronique Mbeng Penlap Leopold Djomkam Tietcheu Damian Nota Anong Akindeh Mbuh Nji Vincent P. K. Titanji Department of Biochemistry and Microbiology Faculty of Science University of Buea Buea Cameroon Laboratory for Public Health Research Biotechnologies Biotechnology Centre University of Yaoundé Yaoundé Cameroon Mezam Polyclinic Bamenda Cameroon

Drug sensitivity testing to establish resistance to TB drugs takes many months to arrive at. Public health physicians have difficulties with such an approach due to long wait periods and cannot use it to establish community wide prevalence as a way to understand where resistance may be emerging faster and to limit its spread. The objective of this study was to use the dot-blot hybridization technique in the detection of resistance to rifamycin (RIF) and streptomycin (SM) in South- Western Cameroon and to compare the technique with the routine culture and drug susceptibility testing for detecting resistance in a resource poor country, Cameroon. A hospital-based study was conducted at the Regional hospitals of Buea and Limbe and Tiko Central Clinic. Tuberculosis (TB) patients aged 15 to 50 (mean age: 30.50 ± 8.33 standard deviation) were recruited for the study between December 2006 and April 2007. Cultures from 59 patients were tested for rifampicin and streptomycin sensitivity by the modified proportion method and mutational analysis for rpoB codon 516 and rrs codon 513 was performed by the dot-blot hybridization technique. Of the 59 sputum samples collected (36 were males and 23 were females) came from Buea 19 (32.2%), Limbe 20 (33.9%) and Tiko 20 (33.9%) towns respectively. Amplification for the gene showed that there was (59) 100% amplification with primers used for rpoB genes and 43 (72.9%) amplification with primers used for the rrs gene. Mutational analysis demonstrated that resistance to RIF was common in females (52.1%) than males (41.7%) while 6% of the samples were indeterminate. 12 (20.3%) samples showed phenotypic and genotypic resistance to RIF compared to 34 samples (58.1%) for SM. Phenotypic resistance and genotypic susceptibility were found in 5 (8.5%) RIF and 3 (4.7%) SM compared to phenotypic susceptibility and genotypic resistance that were found in 2 (3.5%) RIF and 3(4.7%) SM. Double mutation on rpoB and rrs genes occurred in 8 (13.6%) DNA sample

关键词： pcr-based dot-blot analysis Rifamycin Streptomycin SW Region

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