OBJECTIVES: To study the suitability, stability and diversity of short tandem repeat (STR) genomic markers to elicit strain variation in the Mycobacterium leprae isolates within leprosy patients from Andhra Pradesh and Tamil Nadu states in South India. MATERIALS AND METHODS: Slit skin smear (SSS) samples were collected from lesions and various body sites of newly diagnosed leprosy patients. The SSSs from each patient were pooled, except in the case of five patients. Total DNA was extracted from SSS samples. M. leprae STRs were amplified from the DNA either by multiplex PCR (MP) or single PCR methods. The number of repeats for each STR locus (the STR allele) was obtained either by fragment length analysis (FLA) or by DNA sequencing of the PCR amplicons. RESULTS AND CONCLUSION: Multiplex PCR minimised the use of DNA and reagents, and together with FLA, was time and cost effective for STR strain typing. After examination of the isolates of South Indian origin at 13 STR loci, it was determined that the alleles for (AC)8b, (GGT)5, 6-3a (rpoT), 21-3, 27-5, and 23-3 were conserved in two study populations. In a family from Andhra Pradesh, the M. leprae STR patterns in two patients were identical in 16 of 18 loci which indicate a common source of infection. Fourteen of 15 STR loci showed no intra-patient variation in the five patients tested in Tamil Nadu. Altogether, these studies indicate the suitability of STR strain typing for assessing short-range transmission chains.
OBJECTIVES: To study the suitability, stability and diversity of short tandem repeat (STR) genomic markers to elicit strain variation in the Mycobacterium leprae isolates within leprosypatients from Andhra Pradesh and Tamil Nadu states in South India. MATERIALS AND METHODS: Slit skin smear (SSS) samples were collected from lesions and various body sites of newly diagnosed leprosypatients. The SSSs from each patient were pooled, except in the case of five patients. Total DNA was extracted from SSS samples. M. leprae STRs were amplified from the DNA either by multiplex PCR (MP) or single PCR methods. The number of repeats for each STR locus (the STR allele) was obtained either by fragment length analysis (FLA) or by DNA sequencing of the PCR amplicons. RESULTS AND CONCLUSION: Multiplex PCR minimised the use of DNA and reagents, and together with FLA, was time and cost effective for STR strain typing. After examination of the isolates of South Indian origin at 13 STR loci, it was determined that the alleles for (AC)8b, (GGT)5, 6-3a (rpoT), 21-3, 27-5, and 23-3 were conserved in two study populations. In a family from Andhra Pradesh, the M. leprae STR patterns in two patients were identical in 16 of 18 loci which indicate a common source of infection. Fourteen of 15 STR loci showed no intra-patient variation in the five patients tested in Tamil Nadu. Altogether, these studies indicate the suitability of STR strain typing for assessing short-range transmission chains.
Authors: Luana Nepomueceno Costa Lima; Cristiane Cunha Frota; Phillip Noel Suffys; Amanda Nogueira Brum Fontes; Rosa Maria Salani Mota; Rosa Livia Freitas Almeida; Maria Araci de Andrade Pontes; Heitor de Sá Gonçalves; Carl Kendall; Ligia Regina Sansigolo Kerr Journal: Pathog Glob Health Date: 2018-02-06 Impact factor: 2.894
Authors: Wei Li; Rama M Sakamuri; Danielle E Lyons; Florenda M Orcullo; Vidyagouri Shinde; Edred Lao Dela Pena; Armi A Maghanoy; Irene B Mallari; Esterlina V Tan; Indira Nath; Patrick J Brennan; Marivic Balagon; Varalakshmi Vissa Journal: Antimicrob Agents Chemother Date: 2011-08-22 Impact factor: 5.191
Authors: Amanda N B Fontes; Luana N G C Lima; Rosa M S Mota; Rosa L F Almeida; Maria A Pontes; Heitor de S Gonçalves; Cristiane C Frota; Varalakshmi D Vissa; Patrick J Brennan; Ricardo J P S Guimaraes; Carl Kendall; Ligia R F S Kerr; Philip N Suffys Journal: PLoS Negl Trop Dis Date: 2017-12-15