{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T15:05:26Z","timestamp":1790867126132,"version":"4.1.0"},"reference-count":25,"publisher":"Ovid Technologies (Wolters Kluwer Health)","issue":"1","content-domain":{"domain":["lww.com","ovid.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2017,1]]},"abstract":"\n Objective\/Background:<\/jats:title>\n \n Clinical diagnosis of indeterminate and tuberculoid leprosy is often difficult due to limited and confounding signs and symptoms. In the current study, we evaluated the utility of new multiplex polymerase chain reaction (PCR) using\n Mycobacterium leprae<\/jats:italic>\n -specific DNA sequences in the pseudogene regions of\n ML1545<\/jats:italic>\n ,\n ML2180<\/jats:italic>\n , and\n ML2179<\/jats:italic>\n for PCR-based diagnosis of indeterminate leprosy (IND) and leprosy cases across the immunological spectrum. The sensitivity was compared with that of\n RLEP<\/jats:italic>\n PCR.\n <\/jats:p>\n <\/jats:sec>\n \n Methods:<\/jats:title>\n \n DNA was extracted from paraffin-embedded skin biopsy specimens of 220 leprosy cases, which were divided into IND (41), tuberculoid form (3), borderline tuberculoid (42), midborderline (3), borderline lepromatous (\n n<\/jats:italic>\n =59), and lepromatous leprosy (72) cases. PCR positivity of both multiplex and\n RLEP<\/jats:italic>\n PCR were compared in all the samples. A decision tree was constructed using the classification and regression trees algorithm to predict the probability of PCR positivity with the new multiplex PCR scheme in various clinical groups of leprosy. Sensitivity of each pseudogene target was determined using real-time PCR assays, and specificity was confirmed by PCR amplification of DNA extracted from three other mycobacterial species and skin biopsies of 44 non-leprosy cases.\n <\/jats:p>\n <\/jats:sec>\n \n Results:<\/jats:title>\n \n A multiplex PCR positivity of 75.61% was noted in IND cases when compared to that of 58.54% using RLEP PCR (\n P<\/jats:italic>\n < 0.05). Enhanced multiplex PCR positivity was noted across various clinical groups in comparison to\n RLEP<\/jats:italic>\n PCR. The decision tree classifier has predicted statistically significant probability for multiplex PCR positivity among\n RLEP<\/jats:italic>\n -PCR negative group and clinical groups with a low bacillary load.\n <\/jats:p>\n <\/jats:sec>\n \n Conclusion:<\/jats:title>\n This new multiplex PCR scheme can support the diagnosis of indeterminate and tuberculoid forms of leprosy with limited clinical manifestations and can be implemented in basic clinical\/diagnostic setting that possess conventional PCR facilities.<\/jats:p>\n <\/jats:sec>","DOI":"10.4103\/2212-5531.201885","type":"journal-article","created":{"date-parts":[[2017,5,18]],"date-time":"2017-05-18T11:07:10Z","timestamp":1495105630000},"page":"1-8","update-policy":"https:\/\/doi.org\/10.1097\/lww.0000000000001000","source":"Crossref","is-referenced-by-count":24,"title":["Analysis of a Novel Multiplex Polymerase Chain Reaction Assay as a Sensitive Tool for the Diagnosis of Indeterminate and Tuberculoid Forms of Leprosy"],"prefix":"10.4103","volume":"6","author":[{"given":"V. Sundeep","family":"Chaitanya","sequence":"first","affiliation":[{"name":"Division of Molecular Biology and Immunology, Schieffelin Institute of Health Research and Leprosy Centre, Karigiri, Vellore, Tamil Nadu, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luz","family":"Cuello","sequence":"additional","affiliation":[{"name":"Department of Biology, Regents Hall of Natural Science, Northfield, MN 55057, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Madhusmita","family":"Das","sequence":"additional","affiliation":[{"name":"Division of Molecular Biology and Immunology, Schieffelin Institute of Health Research and Leprosy Centre, Karigiri, Vellore, Tamil Nadu, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"A.","family":"Sudharsan","sequence":"additional","affiliation":[{"name":"Division of Molecular Biology and Immunology, Schieffelin Institute of Health Research and Leprosy Centre, Karigiri, Vellore, Tamil Nadu, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pavithrra","family":"Ganesan","sequence":"additional","affiliation":[{"name":"Division of Molecular Biology and Immunology, Schieffelin Institute of Health Research and Leprosy Centre, Karigiri, Vellore, Tamil Nadu, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"K.","family":"Kanmani","sequence":"additional","affiliation":[{"name":"Division of Molecular Biology and Immunology, Schieffelin Institute of Health Research and Leprosy Centre, Karigiri, Vellore, Tamil Nadu, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lakshmi","family":"Rajan","sequence":"additional","affiliation":[{"name":"Division of Molecular Biology and Immunology, Schieffelin Institute of Health Research and Leprosy Centre, Karigiri, Vellore, Tamil Nadu, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mannam","family":"Ebenezer","sequence":"additional","affiliation":[{"name":"Division of Molecular Biology and Immunology, Schieffelin Institute of Health Research and Leprosy Centre, Karigiri, Vellore, Tamil Nadu, India"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"276","reference":[{"key":"R1-20250603","doi-asserted-by":"crossref","first-page":"1096","DOI":"10.1086\/512809","article-title":"Methods for the classification of leprosy for treatment purposes","volume":"44","author":"Pardillo","year":"2007","journal-title":"Clin Infect Dis"},{"key":"R2-20250603","doi-asserted-by":"crossref","first-page":"158","DOI":"10.4103\/0019-5154.127676","article-title":"Clinical profile of leprosy patients: A prospective study","volume":"59","author":"Thakkar","year":"2014","journal-title":"Indian J Dermatol"},{"key":"R3-20250603","first-page":"172","article-title":"Histology of indeterminate leprosy","volume":"50","author":"Liu","year":"1982","journal-title":"Int J Lepr Other Mycobact Dis"},{"key":"R4-20250603","doi-asserted-by":"crossref","first-page":"409","DOI":"10.1002\/mus.20465","article-title":"Pure neural leprosy: Diagnostic value of the polymerase chain reaction","volume":"33","author":"Bezerra Da Cunha","year":"2006","journal-title":"Muscle Nerve"},{"key":"R5-20250603","doi-asserted-by":"crossref","first-page":"41","DOI":"10.47276\/lr.84.1.41","article-title":"Pure neuritic leprosy in patients from a high endemic region of Colombia","volume":"84","author":"Rodriguez","year":"2013","journal-title":"Lepr Rev"},{"key":"R6-20250603","doi-asserted-by":"crossref","first-page":"e2655","DOI":"10.1371\/journal.pntd.0002655","article-title":"PCR-based techniques for leprosy diagnosis: From the laboratory to the clinic","volume":"8","author":"Martinez","year":"2014","journal-title":"PLoS Negl Trop Dis"},{"key":"R7-20250603","doi-asserted-by":"crossref","first-page":"252","DOI":"10.1186\/1471-2334-10-252","article-title":"Multiplex PCR technique could be an alternative approach for early detection of leprosy among close contacts \u2013 A pilot study from India","volume":"10","author":"Banerjee","year":"2010","journal-title":"BMC Infect Dis"},{"key":"R8-20250603","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1016\/j.proche.2016.01.036","article-title":"Detection of Mycobacterium leprae in formalin-fixed paraffin-embedded sample by Fite-Faraco staining and polymerase chain reaction","volume":"18","author":"Sandhika","year":"2016","journal-title":"Procedia Chem"},{"key":"R9-20250603","doi-asserted-by":"crossref","first-page":"906","DOI":"10.1128\/jcm.29.5.906-910.1991","article-title":"Application of a polymerase chain reaction for the detection of Mycobacterium leprae in skin tissues","volume":"29","author":"de Wit","year":"1991","journal-title":"J Clin Microbiol"},{"key":"R10-20250603","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1016\/j.ijmyco.2016.01.002","article-title":"Mycobacterium leprae specific genomic target in the promoter region of probable 4-alpha-glucanotransferase (ML1545) gene with potential sensitivity for polymerase chain reaction based diagnosis of leprosy","volume":"5","author":"Sundeep Chaitanya","year":"2016","journal-title":"Int J Mycobacteriol"},{"key":"R11-20250603","doi-asserted-by":"crossref","first-page":"O142","DOI":"10.1111\/1469-0691.12365","article-title":"Single nucleotide polymorphism-based molecular typing of M. leprae from multicase families of leprosy patients and their surroundings to understand the transmission of leprosy","volume":"20","author":"Turankar","year":"2014","journal-title":"Clin Microbiol Infect"},{"key":"R12-20250603","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1007\/s00430-014-0354-1","article-title":"A report of rifampin-resistant leprosy from northern and eastern India: Identification and in silico analysis of molecular interactions","volume":"204","author":"Vedithi","year":"2015","journal-title":"Med Microbiol Immunol"},{"key":"R13-20250603","doi-asserted-by":"crossref","first-page":"524","DOI":"10.4269\/ajtmh.13-0659","article-title":"Application of RLEP real-time PCR for detection of M. leprae DNA in paraffin-embedded skin biopsy specimens for diagnosis of paucibacillary leprosy","volume":"90","author":"Yan","year":"2014","journal-title":"Am J Trop Med Hyg"},{"key":"R14-20250603","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.ins.2013.12.060","article-title":"The CART decision tree for mining data streams","volume":"266","author":"Rutkowski","year":"2014","journal-title":"Information Sciences"},{"key":"R18-20250603","doi-asserted-by":"crossref","first-page":"e328","DOI":"10.1371\/journal.pntd.0000328","article-title":"Enumeration of Mycobacterium leprae using real-time PCR","volume":"2","author":"Truman","year":"2008","journal-title":"PLoS Negl Trop Dis"},{"key":"R19-20250603","first-page":"199","article-title":"Histopathological study of early skin lesions in leprosy","volume":"94","author":"Nayar","year":"1972","journal-title":"Arch Pathol"},{"key":"R20-20250603","doi-asserted-by":"crossref","first-page":"191","DOI":"10.1002\/path.1711110307","article-title":"The pathogenesis of the early skin lesion in leprosy","volume":"111","author":"Ridley","year":"1973","journal-title":"J Pathol"},{"key":"R21-20250603","first-page":"257","article-title":"The use of histopathology in leprosy diagnosis and research","volume":"60","author":"Lucas","year":"1989","journal-title":"Lepr Rev"},{"key":"R22-20250603","first-page":"12","article-title":"Indeterminate leprosy: Histopathologic and histochemical predictive parameters involved in its possible change to paucibacillary or multibacillary leprosy","volume":"59","author":"Takahashi","year":"1991","journal-title":"Int J Lepr Other Mycobact Dis"},{"key":"R23-20250603","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1128\/jcm.33.1.45-49.1995","article-title":"Detection of Mycobacterium leprae infection by PCR","volume":"33","author":"Wichitwechkarn","year":"1995","journal-title":"J Clin Microbiol"},{"key":"R24-20250603","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1590\/S0100-879X2012007500011","article-title":"Use of the polymerase chain reaction to detect Mycobacterium leprae in urine","volume":"45","author":"Caleffi","year":"2012","journal-title":"Braz J Med Biol Res"},{"key":"R25-20250603","doi-asserted-by":"crossref","first-page":"54","DOI":"10.1016\/j.ijmyco.2014.11.062","article-title":"Comparative evaluation of PCR amplification of RLEP, 16S rRNA, rpoT and Sod A gene targets for detection of M. leprae DNA from clinical and environmental samples","volume":"4","author":"Turankar","year":"2015","journal-title":"Int J Mycobacteriol"},{"key":"R26-20250603","doi-asserted-by":"crossref","first-page":"693","DOI":"10.4103\/0378-6323.116740","article-title":"Fite-Faraco staining in combination with multiplex polymerase chain reaction: A new approach to leprosy diagnosis","volume":"79","author":"Reja","year":"2013","journal-title":"Indian J Dermatol Venereol Leprol"},{"key":"R27-20250603","doi-asserted-by":"crossref","first-page":"e196","DOI":"10.1371\/journal.pntd.0000196","article-title":"Decision tree algorithms predict the diagnosis and outcome of dengue fever in the early phase of illness","volume":"2","author":"Tanner","year":"2008","journal-title":"PLoS Negl Trop Dis"},{"key":"R29-20250603","doi-asserted-by":"crossref","first-page":"e1354","DOI":"10.1371\/journal.pntd.0001354","article-title":"Evaluation of qPCR-based assays for leprosy diagnosis directly in clinical specimens","volume":"5","author":"Martinez","year":"2011","journal-title":"PLoS Negl Trop Dis"}],"container-title":["The International Journal of Mycobacteriology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.ovid.com\/01929415-201706010-00001","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,27]],"date-time":"2026-08-27T13:19:00Z","timestamp":1787836740000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.ovid.com\/01929415-201706010-00001"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,1]]},"references-count":25,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2017]]}},"URL":"https:\/\/doi.org\/10.4103\/2212-5531.201885","relation":{},"ISSN":["2212-5531","2212-554X"],"issn-type":[{"value":"2212-5531","type":"print"},{"value":"2212-554X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2017,1]]}}}