phosphorus-radioisotopes has been researched along with Tuberculosis--Multidrug-Resistant* in 2 studies
2 other study(ies) available for phosphorus-radioisotopes and Tuberculosis--Multidrug-Resistant
| Article | Year |
|---|---|
Detection of multidrug-resistant Mycobacterium tuberculosis directly from sputum samples of patients from Jakarta, Indonesia by radioisotope-based PCR-dot blot hybridization.
The problem of eradicating tuberculosis (TB) has become more complicated by the emergence of multidrug resistant TB (MDR-TB). Any rapid laboratory method that can be used to detect drug susceptibility of Mycobacterium tuberculosis (MTB) is urgently needed. In this study, we employed the radioisotope (32P)-based PCR-dot blot hybridization method on sputum samples from patients in Jakarta, Indonesia. Bacterial DNA was extracted using BOOM method. KatG and rpobeta were amplified by PCR and katG315 or rpobeta531 mutations were identified by dot blot hybridization. Of 100 samples, 11% and 22% showed presence of mutation at codons 315 (AGC --> ACC) of katG and 531 (TCG --> TTG) of rpobeta, respectively. Five percent of the samples showed both mutations. This method is rapid, sensitive, and reliable and can be used to screen large numbers of samples in epidemiological studies. Topics: Antitubercular Agents; Bacterial Proteins; Catalase; Codon; DNA-Directed RNA Polymerases; DNA, Bacterial; Female; Humans; Immunoblotting; Indonesia; Male; Mutation; Nucleic Acid Hybridization; Phosphorus Radioisotopes; Polymerase Chain Reaction; Sensitivity and Specificity; Sputum; Tuberculosis, Multidrug-Resistant | 2012 |
Rapid detection of ethambutol-resistant Mycobacterium tuberculosis directly from sputum samples by radioisotope (32P)-based PCR dot blot hybridization and sequencing methods.
to develop an assay system of a radioisotope (32P)-based PCR dot-blot hybridization technique and evaluation of the assay directly for TB sputum samples to detect mutation at codon 306 of embB gene of Mycobacterium tuberculosis related with ethambutol (EMB) resistance.. one hundred and sixteen of sputum samples were used in this study. Bacterial genome in sputum samples was extracted and tested for mutation at codon 306 of embB gene by the developed PCR dot blot assay using a radioisotope (32P)-labeled oligonucleotide. The positive results were confirmed by DNA sequencing.. all 116 sputum samples were PCR positive for M. tuberculosis. Of 116 samples, three (2.59%) were EMB resistant-M. tuberculosis (MTB) and showed a substitution mutation (ATG/Met'-->GTG/Val) at codon 306 of embB gene. None of mutation was detected at codon 299 of embB gene.. we successfully developed a radioisotope (32P)-based PCR dot blot hybridization technique for detection of mutation at codon 306 of embB gene related with EMB resistant M. tuberculosis. The assay can detect a large number of samples that is suitable for monitoring, surveillance, and epidemiology studies. Topics: Antitubercular Agents; Base Sequence; Codon; Drug Resistance, Multiple, Bacterial; Ethambutol; Feasibility Studies; Genotype; Humans; Mutation; Mycobacterium tuberculosis; Nucleic Acid Hybridization; Pentosyltransferases; Phosphorus Radioisotopes; Polymerase Chain Reaction; Sputum; Time Factors; Tuberculosis, Multidrug-Resistant | 2011 |