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Low frequency of moaA3 gene among the clinical isolates of Mycobacterium tuberculosis from Tamil Nadu and Pondicherry--south eastern coastal states of India.

Sekar B, Arunagiri K, Selvakumar N, Preethi KS, Menaka K - BMC Infect. Dis. (2009)

Bottom Line: In BCG it was absent as expected, but a 386 bp fragment was amplified.The second PCR amplified the flanking sequence of moaA3 and yielded the expected amplicon of 1254 bp in all those 10.3% of clinical isolates which had the 386 bp fragment.However the earlier study carried out in Kerala, reported the presence of moaA3 gene in majority (97%) of their clinical isolates.

View Article: PubMed Central - HTML - PubMed

Affiliation: Division of Laboratories, Central Leprosy Teaching and Research Institute, Chengalpattu, Tamil Nadu, India. drbsekar@yahoo.com

ABSTRACT

Background: Comparative genomic analysis of M. tuberculosis H37Rv and M. bovis BCG have shown that 16 RDs (Regions of Differences) are deleted in BCG and have shown six deletion regions in M. tuberculosis H37Rv. RD1, is present in M. tuberculosis but is absent in all M. bovis BCG sub-strains. A study from Kerala, a south-western coastal state of India aimed to find out differences in RD1 region showed for the first time the presence of moaA3 gene in majority of their clinical isolates, that was absent in type strain H37Rv. We attempted to find out such polymorphism between type strains and the clinical isolates within RD1, targeting moaA3 gene among the clinical isolates of Tamil Nadu & Pondicherry, south-eastern coastal states of India

Methods: One hundred and sixteen clinical isolates of M. tuberculosis were included in the study. PCR using RD1DLa and RD1DRa primers was carried out to amplify a 652 bp fragment, encoding for cfp10 and esat 6 proteins of RD1. A second PCR using primers designed from the surrounding regions of moaA3 gene was done to confirm the presence of the full Open Reading Frame (ORF) in clinical isolates.

Results: In M. tuberculosis H37Rv the expected 652 bp band was present. In BCG it was absent as expected, but a 386 bp fragment was amplified. Around 12/116 (10.3%) of our clinical isolates showed both 652 and 386 bp fragments. The additional 386 bp amplicon is a part of the moaA3 gene which codes for molybdopterin cofactor protein A in M. bovis. The second PCR amplified the flanking sequence of moaA3 and yielded the expected amplicon of 1254 bp in all those 10.3% of clinical isolates which had the 386 bp fragment. However the earlier study carried out in Kerala, reported the presence of moaA3 gene in majority (97%) of their clinical isolates.

Conclusion: This finding showed that there was regional variation presenting polymorphism in moA3 gene, among the strains of M. tuberculosis and further strengthens the speculation of genetic differences among the strains of Kerala and Tamil Nadu & Pondicherry, the South Indian states.

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PCR for amplification of flanking sequence of 386 bp of moaA3 gene. Lane-1 – Molecular weight marker (500 bp ladder). Lane-2 to 7 – Clinical isolates which were positive for 386 bp extra amplicon, showing 1254 bp of flanking sequence. Lane-8 – Negative control.
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Figure 2: PCR for amplification of flanking sequence of 386 bp of moaA3 gene. Lane-1 – Molecular weight marker (500 bp ladder). Lane-2 to 7 – Clinical isolates which were positive for 386 bp extra amplicon, showing 1254 bp of flanking sequence. Lane-8 – Negative control.

Mentions: The second PCR using primers designed from the surrounding region of moaA3 gene done to confirm the presence of the full ORF in clinical isolates, amplified the expected 1254 bp in all the clinical isolates which showed the extra 386 bp amplicon. (Figure 2) (Table 2)


Low frequency of moaA3 gene among the clinical isolates of Mycobacterium tuberculosis from Tamil Nadu and Pondicherry--south eastern coastal states of India.

Sekar B, Arunagiri K, Selvakumar N, Preethi KS, Menaka K - BMC Infect. Dis. (2009)

PCR for amplification of flanking sequence of 386 bp of moaA3 gene. Lane-1 – Molecular weight marker (500 bp ladder). Lane-2 to 7 – Clinical isolates which were positive for 386 bp extra amplicon, showing 1254 bp of flanking sequence. Lane-8 – Negative control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2721839&req=5

Figure 2: PCR for amplification of flanking sequence of 386 bp of moaA3 gene. Lane-1 – Molecular weight marker (500 bp ladder). Lane-2 to 7 – Clinical isolates which were positive for 386 bp extra amplicon, showing 1254 bp of flanking sequence. Lane-8 – Negative control.
Mentions: The second PCR using primers designed from the surrounding region of moaA3 gene done to confirm the presence of the full ORF in clinical isolates, amplified the expected 1254 bp in all the clinical isolates which showed the extra 386 bp amplicon. (Figure 2) (Table 2)

Bottom Line: In BCG it was absent as expected, but a 386 bp fragment was amplified.The second PCR amplified the flanking sequence of moaA3 and yielded the expected amplicon of 1254 bp in all those 10.3% of clinical isolates which had the 386 bp fragment.However the earlier study carried out in Kerala, reported the presence of moaA3 gene in majority (97%) of their clinical isolates.

View Article: PubMed Central - HTML - PubMed

Affiliation: Division of Laboratories, Central Leprosy Teaching and Research Institute, Chengalpattu, Tamil Nadu, India. drbsekar@yahoo.com

ABSTRACT

Background: Comparative genomic analysis of M. tuberculosis H37Rv and M. bovis BCG have shown that 16 RDs (Regions of Differences) are deleted in BCG and have shown six deletion regions in M. tuberculosis H37Rv. RD1, is present in M. tuberculosis but is absent in all M. bovis BCG sub-strains. A study from Kerala, a south-western coastal state of India aimed to find out differences in RD1 region showed for the first time the presence of moaA3 gene in majority of their clinical isolates, that was absent in type strain H37Rv. We attempted to find out such polymorphism between type strains and the clinical isolates within RD1, targeting moaA3 gene among the clinical isolates of Tamil Nadu & Pondicherry, south-eastern coastal states of India

Methods: One hundred and sixteen clinical isolates of M. tuberculosis were included in the study. PCR using RD1DLa and RD1DRa primers was carried out to amplify a 652 bp fragment, encoding for cfp10 and esat 6 proteins of RD1. A second PCR using primers designed from the surrounding regions of moaA3 gene was done to confirm the presence of the full Open Reading Frame (ORF) in clinical isolates.

Results: In M. tuberculosis H37Rv the expected 652 bp band was present. In BCG it was absent as expected, but a 386 bp fragment was amplified. Around 12/116 (10.3%) of our clinical isolates showed both 652 and 386 bp fragments. The additional 386 bp amplicon is a part of the moaA3 gene which codes for molybdopterin cofactor protein A in M. bovis. The second PCR amplified the flanking sequence of moaA3 and yielded the expected amplicon of 1254 bp in all those 10.3% of clinical isolates which had the 386 bp fragment. However the earlier study carried out in Kerala, reported the presence of moaA3 gene in majority (97%) of their clinical isolates.

Conclusion: This finding showed that there was regional variation presenting polymorphism in moA3 gene, among the strains of M. tuberculosis and further strengthens the speculation of genetic differences among the strains of Kerala and Tamil Nadu & Pondicherry, the South Indian states.

Show MeSH
Related in: MedlinePlus