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Journal of Clinical Microbiology, September 2009, p. 2844-2854, Vol. 47, No. 9
0095-1137/09/$08.00+0     doi:10.1128/JCM.02021-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Population-Based Molecular Epidemiology of Leprosy in Cebu, Philippines {triangledown}

Rama Murthy Sakamuri,1 Miyako Kimura,1 Wei Li,1 Hyun-Chul Kim,2 Hyeyoung Lee,2 Madanahally D. Kiran,1 William C. Black IV,1 Marivic Balagon,3 Robert Gelber,3 Sang-Nae Cho,2 Patrick J. Brennan,1 and Varalakshmi Vissa1*

Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado,1 Department of Microbiology, College of Medicine, Yonsei University, Seoul, Republic of Korea,2 Leonard Wood Memorial, Cebu Skin Clinic, Cebu, Philippines3

Received 18 October 2008/ Returned for modification 22 May 2009/ Accepted 8 June 2009

To address the persisting problem of leprosy in Cebu, Philippines, we compiled a database of more than 200 patients who attend an established referral skin clinic. We described the patient characteristics in conventional demographic parameters and also applied multiple-locus variable-number tandem-repeat (VNTR) analysis (MLVA) and single nucleotide polymorphism (SNP) typing for Mycobacterium leprae in biopsied skin lesion samples. These combined approaches revealed that transmission is ongoing, with the affected including the young Cebuano population under 40 years of age in both crowded cities and rural areas of the island. The emergence of multicase families (MCF) is indicative of infection unconstrained by standard care measures. For the SNPs, we designed a low-cost PCR-restriction fragment length polymorphism typing method. MLVA in M. leprae was highly discriminatory in this population yet could retain broad groups, as defined by the more stable SNPs, implying temporal marker stability suitable for interpreting population structures and evolution. The majority of isolates belong to an Asian lineage (SNP type 1), and the rest belong to a putative postcolonial lineage (SNP type 3). Specific alleles at two VNTR loci, (GGT)5 and 21-3, were highly associated with SNP type 3 in this population. MLVA identified M. leprae genotype associations for patients with known epidemiological links such as in MCFs and in some villages. These methods provide a molecular database and a rational framework for targeted approaches to search and confirm leprosy transmission in various scenarios.

* Corresponding author. Mailing address: Department of Microbiology, Immunology, and Pathology, Colorado State University, 1682 Campus Delivery, Fort Collins, CO 80523. Phone: (970) 491-0752. Fax: (970) 491-1815. E-mail: vvissa{at}colostate.edu

{triangledown} Published ahead of print on 1 July 2009.

Journal of Clinical Microbiology, September 2009, p. 2844-2854, Vol. 47, No. 9
0095-1137/09/$08.00+0     doi:10.1128/JCM.02021-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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