03353nas a2200301   4500000000100000008004100001260001200042653001400054653000800068653001200076653003100088653002800119100001200147700001300159700001400172700001200186700001300198700001300211700001100224700001500235700001200250700001500262245008900277856026000366300000900626520240200635022001403037       2023                            d  c12/202310adiagnosis10aENL10aleprosy10aMycobacterium lepromatosis10aWhole genome sequencing1 aSingh I1 aPathak V1 aLavania M1 aAhuja M1 aSharma R1 aNarang T1 aJain S1 aTurankar R1 aDogra S1 aSengupta U00aGenomic characterization of Mycobacterium lepromatosis from ENL patients from India.  uhttps://pdf.sciencedirectassets.com/272223/1-s2.0-S1567134823X00094/1-s2.0-S1567134823001351/main.pdf?X-Amz-Security-Token=IQoJb3JpZ2luX2VjELH%2F%2F%2F%2F%2F%2F%2F%2F%2F%2FwEaCXVzLWVhc3QtMSJGMEQCIFy8kyi4cWoAoZSB6C1XaXXfN5iW%2B%2BPQyWwjwBYlT0KCAiAdFLfJ20ho  a1-113 a<p><strong>Background: </strong>Leprosy is caused by Mycobacterium leprae and Mycobacterium lepromatosis. Both organisms cannot be cultured in vitro. M. lepromatosis was found to be associated mainly with diffuse lepromatous leprosy and with Lucio's phenomena initially. Later, M. lepromatosis was observed in borderline leprosy cases (BL), lepromatous leprosy cases (LL) and leprosy reactional cases (T1R and ENL). Although many cases are being reported with similar clinical features like Lucio phenomenon in India but M. lepromatosis was not isolated from these cases. The aim of this study was to screen MB patients and patients with type 2 reaction for the presence of M. lepromatosis.</p>

<p><strong>Methodology: </strong>We recruited a total of 75 multibacillary leprosy cases (45 MB cases without reaction +30 type 2 reaction (ENL) cases) from TLM hospitals Purulia (West Bengal), Barabanki (Uttar Pradesh), Shahdara (Delhi) and PGIMER (Chandigarh), India. Punch biopsies of 5 mm were collected in 70% ethanol from all the study subjects. DNA was extracted followed by Hemi-nested PCR targeting 16S rRNA gene specific for M. lepromatosis. Further, PCR products were processed for Sanger sequencing for an absolute confirmation of M. lepromatosis. Whole genome sequencing was done to confirm the presence of M. lepromatosis.</p>

<p><strong>Result: </strong>We observed presence of M. lepromatosis in 4 necrotic ENL patients by heminested PCR. There was 100% 16S rRNA sequence similarity with M. lepromatosis FJ924 in one case, 98.96% in two cases and in one case it was 90.9% similarity by nucleotide BLAST (BLASTn) by using the NCBI website. On the basis of Sanger sequencing, we noted presence of M. lepromatosis in 3 necrotic ENL patients as one sample only gave 90.9% similarity by BLASTn. On the basis of de novo assembly and genome obtained, only one sample S4 with a 2.9 mb genome size was qualified for downstream analysis. Sixteen M. lepromatosis- specific proteins were identified in this case and the closest species was M. lepromatosis strain FJ924 based on whole genome level phylogeny.</p>

<p><strong>Conclusion: </strong>These results provide valuable insights into the prevalence of M. lepromatosis in ENL patients in different regions of India and contribute to our understanding of the genetic characteristics of this pathogen in the context of leprosy.</p>
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