Identification of drug-resistantMycobacterium tuberculosis by single strand conformation polymorphism and cleavase fragment length polymorphism |
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Authors: | Manju Sharma Manuel Altamirano |
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Institution: | (1) Vancouver General Hospital and Health Sciences, Vancouver, B.C., Canada;(2) 3/7 J, Medical Enclave, 124001 Rohtak, India |
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Abstract: | Twenty isolates ofMycobacterium tuberculosis resistant to rifampicin(RIF), isoniazid(INH) and streptomycin(STR) were analysed by Polymerase Chain Reaction (PCR) amplification
of rpoB, katG and rrs genes to evaluate comparative diagnostic significance of genetic assays. Mutations were identified by
single strand conformation polymorphism (SSCP) and cleavase fragment length polymorphism (CFLP) and were confirmed by DNA
sequencing. SSCP of 4 RIF resistant and 14 INH resistant isolates showed an extra peak at the level of 75-bp and 85-bp respectively,
while 2 STR resistant isolates showed 2 peaks with 9 bases difference. CFLP showed a different pattern among RIF, INH and
STR sensitive and resistant isolates Thus SSCP and CFLP can be used as alternative diagnostic methods for identification of
mutations in RIF, INH and STR resistant strains of M.tuberculosis. |
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Keywords: | Single strand conformation polymorphism Cleavase fragment length polymorphism Mycobacterium tuberculosis Drug resistance |
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