2013
DOI: 10.1128/jcm.03067-12
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Development of a Novel PCR Restriction Analysis of thehsp65Gene as a Rapid Method To Screen for the Mycobacterium tuberculosis Complex and Nontuberculous Mycobacteria in High-Burden Countries

Abstract: bThe limitations of conventional methods of identification of Mycobacterium tuberculosis have led to the development of several nucleic acid amplification techniques which have the advantage of being rapid, sensitive, and specific. However, their expense or the need for technical expertise makes it difficult to use them in regions in which tuberculosis is endemic. A novel PCR restriction analysis (PRA) of the hsp65 gene was therefore developed for rapid screening of clinical isolates to identify Mycobacterium … Show more

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Cited by 24 publications
(14 citation statements)
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“…This technique amplifies and analyses DNA sequences from either the hsp65 or rpoB gene. 11,12 While awaiting culture, polymerase chain reaction may improve the accuracy and speed at which the diagnosis can be made. Current American Thoracic Society/Infectious Diseases Society of America guidelines also recommend drug susceptibility testing for macrolides in cases of MAC infection, which was performed in this case.…”
Section: Discussionmentioning
confidence: 99%
“…This technique amplifies and analyses DNA sequences from either the hsp65 or rpoB gene. 11,12 While awaiting culture, polymerase chain reaction may improve the accuracy and speed at which the diagnosis can be made. Current American Thoracic Society/Infectious Diseases Society of America guidelines also recommend drug susceptibility testing for macrolides in cases of MAC infection, which was performed in this case.…”
Section: Discussionmentioning
confidence: 99%
“…Accordingly, in recent decades, other genes such as recA ( 13 ), hsp65 ( 14 , 15 ), and rpoB have been considered as candidates for phylogenetic studies and diagnosis ( 16 , 17 ). From these, rpoB encoding β subunit of RNA polymerase enzyme is thought to be the most appropriate gene for such purposes, especially in bacteria with close relativity ( 18 - 20 ).…”
Section: Introductionmentioning
confidence: 99%
“…[ 12 13 ] Today, molecular approaches like sequencing, INNO-LiPA mycobacteria v2, and AccuProbe are used for precise and rapid identification of the species, however, high cost of these methods and also the requirement for special facilities impose limitations on their application in developing countries. [ 14 15 16 ] Common typing methods like bacteriophage typing and serotyping are replaced by ribotyping, polymerase chain reaction (PCR)-based methods, fingerprinting plasmid, and analysis of restriction fragments of chromosomal DNA by pulse-field gel electrophoresis in the last decades. [ 17 ] Among molecular approaches for typing, analysis PCR-restriction fragment length polymorphism (RFLP) is an accurate and inexpensive method providing diagnosis and typing of species and subspecies of mycobacteria.…”
Section: Introductionmentioning
confidence: 99%
“…[ 17 ] Among molecular approaches for typing, analysis PCR-restriction fragment length polymorphism (RFLP) is an accurate and inexpensive method providing diagnosis and typing of species and subspecies of mycobacteria. [ 14 18 ] In order to typing mycobacteria by PCR-RFLP analysis (PRA) method, different genes such as 16SrRNA , rpoB , dnaj , and hsp65 were targeted. [ 9 14 ] The variable regions of rpoB gene that is encoding the β subunit of RNA polymerase enzyme are proposed as proper gene for phylogenic analysis, determination of inter-species diversity and diagnosis of bacteria specially ones with close relativity.…”
Section: Introductionmentioning
confidence: 99%
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