Population-based molecular epidemiology of leprosy in Cebu, Philippines.
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.
Sakamuri
Rama Murthy
RM
Department of Microbiology, Immunology, and Pathology, Colorado State University, 1682 Campus Delivery, Fort Collins, CO 80523, USA.
Kimura
Miyako
M
Li
Wei
W
Kim
Hyun-Chul
HC
Lee
Hyeyoung
H
Kiran
Madanahally D
MD
Black
William C
WC
Balagon
Marivic
M
Gelber
Robert
R
Cho
Sang-Nae
SN
Brennan
Patrick J
PJ
Vissa
Varalakshmi
V
D000293:Adolescent; D000328:Adult; D000368:Aged; D001706:Biopsy; D002648:Child; D002675:Child, Preschool; D016172:DNA Fingerprinting; D004269:DNA, Bacterial / Q000235:genetics; D005260:Female; D005838:Genotype; D006801:Humans; D007918:Leprosy / Q000453:epidemiology* / Q000382:microbiology* / Q000635:transmission; D008297:Male; D008875:Middle Aged; D018598:Minisatellite Repeats; D017720:Molecular Epidemiology; D009166:Mycobacterium leprae / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D010679:Philippines / Q000453:epidemiology; D020641:Polymorphism, Single Nucleotide; D012424:Rural Population; D012867:Skin / Q000382:microbiology; D014505:Urban Population; D055815:Young Adult
D004269:DNA, Bacterial