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

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.

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Acknowledgments

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.

Funding

This study was funded by the Indian Council of Medical Research (ICMR-Adhoc Project 5/8/3(11)2014-ECD-1).

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Correspondence to Ravindra P. Turankar.

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The study was approved by the Ethical Committee of The Leprosy Mission Trust, India.

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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

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Keywords

  • Viable M. leprae
  • SNP genotype
  • RLEP gene
  • 16S rRNA gene
  • qRT-PCR