Isolation of Mycobacterium lepromatosis and Development of Molecular Diagnostic Assays to Distinguish Mycobacterium leprae and M. lepromatosis.
Mycobacterium leprae
Mycobacterium lepromatosis
leprosy diagnostic assay
real-time PCR
Journal
Clinical infectious diseases : an official publication of the Infectious Diseases Society of America
ISSN: 1537-6591
Titre abrégé: Clin Infect Dis
Pays: United States
ID NLM: 9203213
Informations de publication
Date de publication:
05 11 2020
05 11 2020
Historique:
received:
26
08
2019
accepted:
12
11
2019
pubmed:
17
11
2019
medline:
28
4
2021
entrez:
17
11
2019
Statut:
ppublish
Résumé
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). 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. 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. The RLPM and RLEP assays will aid healthcare providers in the clinical diagnosis and surveillance of leprosy.
Sections du résumé
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.
Identifiants
pubmed: 31732729
pii: 5626481
doi: 10.1093/cid/ciz1121
pmc: PMC8189713
doi:
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
e262-e269Informations de copyright
© Published by Oxford University Press for the Infectious Diseases Society of America 2019.
Références
Int J Lepr Other Mycobact Dis. 2001 Mar;69(1):1-12
pubmed: 11480310
Clin Microbiol Rev. 2010 Jul;23(3):550-76
pubmed: 20610823
J Drugs Dermatol. 2012 Feb;11(2):168-72
pubmed: 22270197
Lepr Rev. 1986;57 Suppl 3:209-19
pubmed: 3553813
Int J Lepr Other Mycobact Dis. 2005 Sep;73(3):169-88
pubmed: 16830639
Am J Clin Pathol. 2008 Dec;130(6):856-64
pubmed: 19019760
J Clin Microbiol. 2018 Feb 22;56(3):
pubmed: 29305545
Proc Natl Acad Sci U S A. 2015 Apr 7;112(14):4459-64
pubmed: 25831531
N Engl J Med. 2011 Apr 28;364(17):1626-33
pubmed: 21524213
Mayo Clin Proc. 2019 Mar;94(3):417-423
pubmed: 30799052
Emerg Infect Dis. 2017 Nov;23(11):1864-1866
pubmed: 29048278
Microbiology (Reading). 1994 Aug;140 ( Pt 8):1983-7
pubmed: 7522845
Clin Microbiol Infect. 2019 Nov;25(11):1315-1327
pubmed: 31158516
Bioinformatics. 2007 May 15;23(10):1289-91
pubmed: 17379693
PLoS Negl Trop Dis. 2008;2(11):e328
pubmed: 18982056
Int J Dermatol. 2012 Aug;51(8):952-9
pubmed: 22788812
J Bacteriol. 2009 Oct;191(19):6067-74
pubmed: 19633074
Emerg Infect Dis. 2015 Dec;21(12):2127-34
pubmed: 26583204
J Drugs Dermatol. 2012 Feb;11(2):229-33
pubmed: 22270208
Am J Clin Pathol. 2014 Oct;142(4):524-32
pubmed: 25239420
Wkly Epidemiol Rec. ;92(35):501-19
pubmed: 28861986
Int J Lepr. 1959 Apr-Jun;27:110-5
pubmed: 13825746
Front Public Health. 2017 May 19;5:109
pubmed: 28580353
Am J Trop Med Hyg. 2014 Mar;90(3):524-9
pubmed: 24493677
J Med Microbiol. 2005 Mar;54(Pt 3):235-242
pubmed: 15713606
PLoS Negl Trop Dis. 2011 Oct;5(10):e1354
pubmed: 22022631
PLoS Negl Trop Dis. 2019 Mar 18;13(3):e0007284
pubmed: 30883558
Am J Trop Med Hyg. 2017 Jan 11;96(1):260
pubmed: 27879462