TY - JOUR
T1 - Isolation of Mycobacterium lepromatosis and Development of Molecular Diagnostic Assays to Distinguish Mycobacterium leprae and M. lepromatosis
AU - Sharma, Rahul
AU - Singh, Pushpendra
AU - McCoy, Rajiv C.
AU - Lenz, Shannon M.
AU - Donovan, Kelly
AU - Ochoa, Maria T.
AU - Estrada-Garcia, Iris
AU - Silva-Miranda, Mayra
AU - Jurado-Santa Cruz, Fermin
AU - Balagon, Marivic F.
AU - Stryjewska, Barbara
AU - Scollard, David M.
AU - Pena, Maria T.
AU - Lahiri, Ramanuj
AU - Williams, Diana L.
AU - Truman, Richard W.
AU - Adams, Linda B.
N1 - Publisher Copyright:
© 2019 Published by Oxford University Press for the Infectious Diseases Society of America 2019.
PY - 2020/10/15
Y1 - 2020/10/15
N2 - Background: Mycobacterium leprae was thought to be the exclusive causative agent of leprosy until Mycobacterium lepromatosis was identified in a rare form of leprosy known as diffuse lepromatous leprosy (DLL). Methods: We isolated M. lepromatosis from a patient with DLL and propagated it in athymic nude mouse footpads. Genomic analysis of this strain (NHDP-385) identified a unique repetitive element, RLPM, on which a specific real-time quantitative polymerase chain reaction assay was developed. The RLPM assay, and a previously developed RLEP quantitative polymerase chain reaction assay for M. leprae, were validated as clinical diagnostic assays according to Clinical Laboratory Improvement Amendments guidelines. We tested DNA from archived histological sections, patient specimens from the United States, Philippines, and Mexico, and US wild armadillos. Results: The limit of detection for the RLEP and RLPM assays is 30 M. leprae per specimen (0.76 bacilli per reaction; coefficient of variation, 0.65%-2.44%) and 122 M. lepromatosis per specimen (3.05 bacilli per reaction; 0.84%-2.9%), respectively. In histological sections (n = 10), 1 lepromatous leprosy (LL), 1 DLL, and 3 Lucio reactions contained M. lepromatosis; 2 LL and 2 Lucio reactions contained M. leprae; and 1 LL reaction contained both species. M. lepromatosis was detected in 3 of 218 US biopsy specimens (1.38%). All Philippines specimens (n = 180) were M. lepromatosis negative and M. leprae positive. Conversely, 15 of 47 Mexican specimens (31.91%) were positive for M. lepromatosis, 19 of 47 (40.43%) were positive for M. leprae, and 2 of 47 (4.26%) contained both organisms. All armadillos were M. lepromatosis negative. Conclusions: The RLPM and RLEP assays will aid healthcare providers in the clinical diagnosis and surveillance of leprosy.
AB - Background: Mycobacterium leprae was thought to be the exclusive causative agent of leprosy until Mycobacterium lepromatosis was identified in a rare form of leprosy known as diffuse lepromatous leprosy (DLL). Methods: We isolated M. lepromatosis from a patient with DLL and propagated it in athymic nude mouse footpads. Genomic analysis of this strain (NHDP-385) identified a unique repetitive element, RLPM, on which a specific real-time quantitative polymerase chain reaction assay was developed. The RLPM assay, and a previously developed RLEP quantitative polymerase chain reaction assay for M. leprae, were validated as clinical diagnostic assays according to Clinical Laboratory Improvement Amendments guidelines. We tested DNA from archived histological sections, patient specimens from the United States, Philippines, and Mexico, and US wild armadillos. Results: The limit of detection for the RLEP and RLPM assays is 30 M. leprae per specimen (0.76 bacilli per reaction; coefficient of variation, 0.65%-2.44%) and 122 M. lepromatosis per specimen (3.05 bacilli per reaction; 0.84%-2.9%), respectively. In histological sections (n = 10), 1 lepromatous leprosy (LL), 1 DLL, and 3 Lucio reactions contained M. lepromatosis; 2 LL and 2 Lucio reactions contained M. leprae; and 1 LL reaction contained both species. M. lepromatosis was detected in 3 of 218 US biopsy specimens (1.38%). All Philippines specimens (n = 180) were M. lepromatosis negative and M. leprae positive. Conversely, 15 of 47 Mexican specimens (31.91%) were positive for M. lepromatosis, 19 of 47 (40.43%) were positive for M. leprae, and 2 of 47 (4.26%) contained both organisms. All armadillos were M. lepromatosis negative. Conclusions: The RLPM and RLEP assays will aid healthcare providers in the clinical diagnosis and surveillance of leprosy.
KW - Mycobacterium leprae
KW - Mycobacterium lepromatosis
KW - leprosy diagnostic assay
KW - real-time PCR
UR - http://www.scopus.com/inward/record.url?scp=85088415893&partnerID=8YFLogxK
U2 - 10.1093/cid/ciz1121
DO - 10.1093/cid/ciz1121
M3 - Artículo
C2 - 31732729
AN - SCOPUS:85088415893
SN - 1058-4838
VL - 71
SP - E262-E269
JO - Clinical Infectious Diseases
JF - Clinical Infectious Diseases
IS - 8
ER -