The potential role of environmental M. leprae in the transmission of leprosy remains unknown. We investigated role of environment as a possible source of viable M. leprae responsible for transmission of leprosy. The samples were collected from 10 multi-case leprosy families comprising, slit skin smear (SSS) from 9 multibacillary (MB), 16 paucibacillary cases (PB), 22 household contacts, and 38 environmental soil samples. The quantum of viable M. leprae was estimated by qRT-PCR using 16S rRNA gene from soil and SSS. Genotypes of M. leprae were determined by gene sequencing. We could observe presence of viable M. leprae in 11 (44%) leprosy cases (M. leprae 16S rRNA gene copies range from 1.78 × 102 to 8.782 × 109) and 4 (18%) household contacts (M. leprae 16S rRNA gene copies range from 2.54 × 103 and 7.47 × 104). Remarkably, presence of viable M. leprae was also noted in 10 (53%) soil samples where in M. leprae 16S rRNA gene copies ranged from 4.36 × 102 to 7.68 × 102. M leprae subtype 1D was noted in most of the leprosy cases their household contacts and in the surrounding soil samples indicating source of infection in household contacts could be from environment or patients. M. leprae 16S rRNA copies were approximately similar in both PB cases and soil samples along with presence of SNP type 1 subtype 1D in both samples indicating source of M. leprae from patients to contacts was either from patients or environment or both.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
World Health Organization (2000) Guide to eliminate leprosy as a public health problem
Dhillon GP (2006) NLEP-current situation and strategy during the 11th plan period (2007–2012). J Indian Med Assoc104:671–672
WHO (2017) Global leprosy update, accelerating reduction of disease burden. Wkly Epidemiol Rec 92:501–520
Rees RJ, McDougall AC (1997) Airborne infection with Mycobacterium leprae in mice. J Med Microbiol 10:63–68
Job CK, Chehl SK, Hastings RC (1994) Transmission of leprosy in nude mice through thorn pricks. Int J Lepr Other Mycobact Dis 62:395–398
Kotteswaran G, Chacko CJ, Job CK (1980) Skin adnexa in leprosy and their role in the dissemination of M. leprae. Lepr India 52:475–481
Matsuoka M, Izumi S, Budiawan T, Nakata N, Saeki K (1999) Mycobacterium leprae DNA in daily using water as a possible source of leprosy infection. Indian J Lepr 71:61–67
Lavania M, Katoch K, Katoch VM et al (2008) Detection of viable Mycobacterium leprae from environmental soil samples: insights into possible sources for transmission of leprosy. Infect Genet Evol 8:627–631
Turankar RP, Lavania M, Singh M et al (2012) Dynamics of Mycobacterium leprae transmission in environmental context. Deciphering the role of environment as a potential reservoir. Infect Genet Evol 12:121–126
Turankar RP, Singh V, Gupta H, Pathak VK, Ahuja M, Singh I et al (2019) Association of non-tuberculous mycobacteria with Mycobacterium leprae in environment of leprosy endemic regions in India. Infect Genet Evol 72:191–198
Singh V, Turankar RP, Goel A (2019) Molecular characterization of environmental mycobacterial species from leprosy endemic tribal regions of North Purulia District. West Bengal. Int J Mycobacteriol 8:381–389
Arraes M, Holanda MV, Lima L, Sabadia J, Duarte CR, Almeida R, Kendall C, Kerr L, Frota CC (2017) Natural environmental water sources in endemic regions of northeastern Brazil are potential reservoirs of viable Mycobacterium leprae. Memorias do Instituto Oswaldo Cruz 112:805–811
Holanda MV, Marques LEC, Macedo MLB, Pontes MAA, Sabadia JAB, Ker LRFS, Almeida RLF, Frota CC (2017) Presence of Mycobacterium leprae genotype 4 in environmental waters in Northeast Brazil. Revista da Sociedade Brasileira de Medicina Tropical 50:216–222
Turankar RP, Lavania M, Singh M, Sengupta U, Siva sai KSR, Jadhav RS (2016) Presence of viable Mycobacterium leprae in environmental specimens around houses of leprosy patients. Ind J Med Microbiol 34: 315–321
Mohanty PS, Naaz F, Katara D, Misba L, Kumar D, Dwivedi DK, Tiwari AK, Chauhan DS, Bansal AK, Tripathy SP, Katoch K (2016) Viability of Mycobacterium leprae in the environment and its role in leprosy dissemination. Ind J Demato venero lepro 82:23–27
Desikan KV, Sreevatsa (1995) Extended studies on the viability of Mycobacterium leprae outside the human body. Lepr Rev 66:284–295
Kurabachew M, Wondimu A, Ryon JJ (1998) Reverse transcription- PCR detection of Mycobacterium leprae in clinical specimens. J Clin Microbiol 36:1352–1356
Martinez AN, Lahiri R, Pittman TL, Scollard D, Truman R, Moraes MO et al (2009) Molecular determination of Mycobacterium leprae viability by use of realtime PCR. J Clin Microbiol 47:2124–2130
D’Abreu PC, Durães SMB, Estrela RR, Baltazar MCNP, Rochael MC (2000) Hanseníasevirchowianadiagnosticadaatravés de examehistopatológico de lesão oral empaciente com pênfigofoliáceo. An Bras Dermatol 75:339–346
Margarido LC, Rivitti EA, Hanseníase (2005) In R focaccia (sci. Ed.). Tratado de infectologia. Atheneu, São Paulo 3: 939–972
Pontes ARB, Almeida MGC, Xavier MB, Quaresma JAS, Yassui EA (2008) Detecção do DNA de Mycobacterium leprae emsecreção nasal. Rev Bras Enferm 61:734–737
Hastings RC, Gillis TP, Krahenbull JL, Franzblau SG (1988) Leprosy. Clin Microbiol Rev 1:330–338
Girdhar BK, Desikan KV (1979) Discharge of Mycobacterium leprae from the mouth in lepromatous le prosy patients. Leprosy Rev 50:45–50
Fine PE, Sterne JA, Ponnighaus JM et al (1997) Household and dwelling contact as risk factors for leprosy in northern Malawi. Am J Epidemiol 146:91–102
Van Beers SM, Hatta M, Klatser PR (1999) Patient contact is the major determinant in incident leprosy: implications for future control. Int J Lepr Other Mycobact Dis 67:119–128
Monot M, Honoré N, Garnier T, Araoz R, Coppée JY, Lacroix C et al (2005) On the origin of leprosy. Science 308:1040–1042
Monot M, Honoré N, Garnier T, Zidane N, Sherafi D, Paniz-Mondolfi A, Matsuoka M, Taylor GM, Donoghue HD, Bouwman A, Mays S, Watson C, Lockwood D, Khamesipour A, Dowlati Y, Jianping S, Rea TH, Vera-Cabrera L, Stefani MM, Banu S, Macdonald M, Sapkota BR, Spencer JS, Thomas J, Harshman K, Singh P, Busso P, Gattiker A, Rougemont J, Brennan PJ, Cole ST (2009) Comparative genomic and phylogeographic analysis of Mycobacterium leprae. Nat Genet 41:1282–1289
Lavania M, Jadhav RS, Turankar RP, Chaitanya VS, Singh M, Sengupta U (2013) Single nucleotide polymorphisms typing of Mycobacterium leprae reveals focal transmission of leprosy in high endemic regions of India. Clin Microbiol Infect 19:1058–1062
Turankar RP, Lavania M, Chaitanya VS, Sengupta U, Darlong J, Darlong F et al (2014) Single nucleotide polymorphism-based molecular typing of M. leprae from multicase families of leprosy patients and their surroundings to understand the transmission of leprosy. Clin Microbiol Infect Dis 3:142–149
Turankar RP, Pandey S, Lavania M, Singh I, Nigam A, Darlong J, Darlong F, Sengupta U (2015) Comparative evaluation of PCR amplification of RLEP, 16S rRNA, rpoT and Sod A gene targets for detection of M. leprae DNA from clinical and environmental samples. Inter J Mycobacreiol 4:54–59
Jadhav RS, Kamble RR, Shinde VS, Edward S, Edward VK (2005) Use of reverse transcription polymerase chain reaction for the detection of Mycobacterium leprae in the slit-skin smears of leprosy patients. Indian J Lepr 77: 116–127
Turankar RP, Lavania M, Singh I, Singh V, Ahuja M, Pathak VK, Ramesh V (2018) Paucibacillary leprosy: reappraisal using Ziehl-Neelsen staining of slit skin smears and 16S rRNA Real Time Polymerase Chain Reaction of nasal swabs. Lepr Rev 89:272–279
Macedo Arraes MLBM, Holanda MV, Lima LNGC, Sabadia JAB, Duarte CR, Almeida RLF, Kendall C, Kerr LRS, Frota CC (2017) Natural environmental water sources in endemic regions of northeastern Brazil are potential reservoirs of viable Mycobacterium leprae. Mem Inst Oswaldo Cruz 112:805–811
Tió-Coma M, Wijnands T, Pierneef L, Schilling AK, Alam K, Roy JC, Faber WR, Menke H, Pieters T, Stevenson K, Richardus JH, Geluk A (2019) Detection of Mycobacterium leprae DNA in soil: multiple needles in the haystack. Sci Rep 9:3165
Lavania M, Turankar RP, Karri S, Chaitanya VS, Sengupta U, Jadhav RS (2013) Cohort study of the seasonal effect on nasal carriage and the presence of Mycobacterium leprae in an endemic area in the general population. Clin Microbiol Infect 19(10):970–974. https://doi.org/10.1111/1469-0691.12087
Turankar RP, Lavania M, Darlong J, Krovvidi S, Sengupta U, Jadhav R (2019) Survival of Mycobacterium leprae and association with Acanthamoeba from environmental samples in the inhabitant areas of active leprosy cases: a cross sectional study from endemic pockets of Purulia, West Bengal. Infect Genet Evol 72
Chae GT, Kim MJ, Kang TJ, Lee SE, Shin Y, Kim JP, Ko YH, Kim SH, Kim NH (2002) DNA-PCR and RT-PCR for the 18-kDa gene of Mycobacterium leprae to assess the efficacy of multi-drug therapy for leprosy. J Med Microbiol 51:417–422
Matsuoka M, Roa RIL, Budiawan T, Kyaw K, Chae GT (2006) Genotypic analysis of Mycobacterium leprae isolates from Japan and other Asian countries reveals a global transmission pattern of leprosy. FEMS Microbiol Letters 261:150–154
We wish to thank Indian Council of Medical Research (ICMR-Adhoc Project 5/8/3(11)2014-ECD-1) for the financial support. We are likewise grateful to Mr. Atul Roy for assisting us in the sample collection. We also thank Superintendent and staff of TLM, Purulia for their help and assistance during the field work. We wish to acknowledge support of The Leprosy Mission, India, to carry out this work. We also acknowledge Dr. Utpal Sengupta Consultant, Dr. Itu Singh, Scientist at SB lab, and Dr. Joydeepa Darlong, Head-knowledge Management TLMTI, India, for their guidance and encouragement.
This study was funded by the Indian Council of Medical Research (ICMR-Adhoc Project 5/8/3(11)2014-ECD-1).
Conflict of interest
The authors declare that they have no conflict of interest.
The study was approved by the Ethical Committee of The Leprosy Mission Trust, India.
A well-written consent was taken from the patients included in the study.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
Singh, V., Turankar, R.P. & Goel, A. Real-time PCR-based quantitation of viable Mycobacterium leprae strain from clinical samples and environmental sources and its genotype in multi-case leprosy families of India. Eur J Clin Microbiol Infect Dis (2020). https://doi.org/10.1007/s10096-020-03958-w
- Viable M. leprae
- SNP genotype
- RLEP gene
- 16S rRNA gene