Real-time PCR-based quantitation of viable Mycobacterium leprae strain from clinical samples and environmental sources a
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ORIGINAL ARTICLE
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 Vikram Singh 1,2
&
Ravindra P. Turankar 1
&
Anjana Goel 2
Received: 30 April 2020 / Accepted: 17 June 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract 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. Keywords Viable M. leprae . SNP genotype . RLEP gene . 16S rRNA gene . qRT-PCR
Introduction Leprosy (also known as Hansen’s disease) is a chronic infectious communicable disease caused by Mycobacterium (M) leprae or M. lepromatosis that has a predilection for the skin and peripheral nerves. World Health Assembly announced leprosy elimination at a global level by the end of 2000 [1] and India achieved elimination at 2005 [2]. However, India is still reporting > 1 lakh (100,000) new cases every year and the annual new case detection rate (ANCDR) remained stabilized over the last 2 decades indicating the continued transmission of infection in leprosy endemic pockets [3].
* Ravindra P. Turankar [email protected] 1
Stanley Browne Laboratory, The Leprosy Mission Community Hospital, Nand Nagari, Delhi 110093, India
2
GLA University, Mathura, Uttar Pradesh 281406, India
Indirectly, animal experiments have provided sufficient proofs [4, 5] in establishing transmission of leprosy in human via droplet infection through discharge from nasal and oral cavity of leprosy patients through sneezing or coughing. M. leprae has also been shown to escape in the environment from the unbroken skin of lepromatous patients through the sweat
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