Author:
ProCite
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Reference Manager
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-
1.
Lastória JC, Morgado de Abreu MAM, 2014. Leprosy: review of the epidemiological, clinical, and etiopathogenic aspects—part 1. An Bras Dermatol 89: 205–218.
-
2.
Rao PN, Suneetha S, 2018. Current situation of leprosy in India and its future implications. Indian Dermatol Online J 9: 83–89.
-
3.
National Leprosy Eradication Programme, 2018. NLEP Annual data (2017–18). Available at: http://nlep.nic.in. Accessed February 22, 2019.
-
4.
World Health Organisation, 2019. Leishmaniasis Facts Sheet. Available at: https://www.who.int/news-room/fact-sheets/detail/leishmaniasis. Accessed April 12, 2019.
-
5.
Mukherjee A, Ramesh V, Mishra RS, 1993. Post kala-azar dermal leishmaniasis: a light and electron microscopic study of 18 cases. J Cutan Pathol 20: 320–325.
-
6.
World Health Organization, 2010. Control of the leishmaniases, World Health Organ Tech Rep Ser 949: 22–26.
-
7.
Trindade MA, Silva LL, Braz LM, Amato VS, Naafs B, Sotto MN, 2015. Post-kala-azar dermal leishmaniasis and leprosy: case report and literature review. BMC Infect Dis 15: 543.
-
8.
Ramesh V, Ramam M, Singh R, Salotra P, 2008.Hypopigmented post-kala-azar dermal leishmaniasis. Int J Dermatol 47: 414–416.
-
9.
Khandpur S, Ramam M, Sharma VK, Salotra P, Singh MK, Malhotra A, 2003. Nerve involvement in Indian post kala-azar dermal leishmaniasis. Acta Derm Venereol 84: 245–246.
-
10.
Verma S, Kumar R, Katara GK, Singh LC, Negi NS, Ramesh V, Salotra P, 2010. Quantification of parasite load in clinical samples of leishmaniasis patients: IL-10 level correlates with parasite load in visceral leishmaniasis. PLoS One 5: e10107.
-
11.
Ghosh P et al. 2018. Evaluation of real-time PCR for diagnosis of post-kala-azar dermal leishmaniasis in endemic Foci of Bangladesh. Open Forum Infect Dis 5: ofy234.
-
12.
Bhargava A, Ramesh V, Verma S, Salotra P, Bala M, 2018. Revisiting the role of the slit-skin smear in the diagnosis of Indian post-kala-azar dermal leishmaniasis. Indian J Dermatol Venereol Leprol 84: 690–695.
-
13.
Azevedo MC, Ramuno NM, Fachin LR, Tassa M, Rosa PS, Belone AF, Diório SM, Soares CT, Garlet GP, Trombone AP, 2017. QPCR detection of Mycobacterium leprae in biopsies and slit skin smear of different leprosy clinical forms. Braz J Infect Dis 21: 71–78.
-
14.
Torres P, Camarena JJ, Gomez JR, Nogueira JM, Gimeno V, Navarro JC, Olmos A, 2003. Comparison of PCR mediated amplification of DNA and the classical methods for detection of Mycobacterium leprae in different types of clinical samples in leprosy patients and contacts. Lepr Rev 74: 18–30.
-
15.
Cheng X et al. 2019. Development and evaluation of a droplet digital PCR assay for the diagnosis of paucibacillary leprosy in skin biopsy specimens. PLoS Negl Trop Dis 13: e0007284.
-
16.
Barbieri RR et al. 2019. Quantitative polymerase chain reaction in paucibacillary leprosy diagnosis: a follow-up study. PLoS Negl Trop Dis 13: e0007147.
-
17.
De Paz HD, Brotons P, Muñoz-Almagro C, 2014. Molecular isothermal techniques for combating infectious diseases: towards low-cost point-of-care diagnostics. Expert Rev Mol Diagn 14: 827–843.
-
18.
Bühlmann A, Pothier JF, Rezzonico F, Smits TH, Andreou M, Boonham N, Duffy B, Frey JE, 2013. Erwiniaamylovora loop-mediated isothermal amplification (LAMP) assay for rapid pathogen detection and on-site diagnosis of fire blight. J Microbiol Methods 92: 332–339.
-
19.
Tomlinson JA, Dickinson MJ, Boonham N, 2010. Detection of Botrytis cinerea by loop-mediated isothermal amplification. Lett Appl Microbiol 51: 650–657.
-
20.
Lucchi NW, Demas A, Narayanan J, Sumari D, Kabanywanyi A, Kachur SP, Barnwell JW, Udhayakumar V, 2010. Real-time fluorescence loop mediated isothermal amplification for the diagnosis of malaria. PLoS One 29: e13733.
-
21.
Chaitanya VS, Cuello L, Das M, Sudharsan A, Ganesan P, Kanmani K, Rajan L, Ebenezer M, 2017. Analysis of a novel multiplex polymerase chain reaction assay as a sensitive tool for the diagnosis of indeterminate and tuberculoid forms of leprosy. Int J Mycobacteriol 6: 1–8.
-
22.
Verma S, Singh R, Sharma V, Bumb RA, Negi NS, Ramesh V, Salotra P, 2017. Development of a rapid loop-mediated isothermal amplification assay for diagnosis and assessment of cure of Leishmania infection. BMC Infect Dis 17: 223.
-
23.
Verma S, Avishek K, Sharma V, Negi NS, Ramesh V, Salotra P, 2013. Application of loop-mediated isothermal amplification assay for the sensitive and rapid diagnosis of visceral leishmaniasis and post-kala-azar dermal leishmaniasis. Diagn Microbiol Infect Dis 75: 390–395.
-
24.
Truman RW, Andrews PK, Robbins NY, Adams LB, Krahenbuhl JL, Gillis TP, 2008. Enumeration of Mycobacterium leprae using real-time PCR. PLoS Negl Trop Dis 2: e328.
-
25.
Singh A, Ramesh V, 2013. Histopathological features in leprosy, post-kala-azar dermal leishmaniasis, and cutaneous leishmaniasis. Indian J Dermatol Venereol Leprol 79: 360–366.
-
26.
Shao Y, Zhu S, Jin C, Chen F, 2011. Development of multiplex loop-mediated isothermal amplification-RFLP (mLAMP-RFLP) to detect Salmonella spp. and Shigella spp. in milk. Int J Food Microbiol 148: 75–79.
-
27.
Jung JH et al. 2015. Combination of multiplex reverse-transcription loop-mediated isothermal amplification with an immunochromatographic strip for subtyping influenza A virus. Anal Chim Acta 853: 541–547.
-
28.
Ball CS, Light YK, Koh CY, Wheeler SS, Coffey LL, Meagher RJ, 2016. Quenching of unincorporated amplification signal reporters in reverse-transcription loop-mediated isothermal amplification enabling bright, single-step, closed-tube, and multiplexed detection of RNA viruses. Anal Chem 88: 3562–3568.
-
29.
Maltempe FG, Baldin VP, Lopes MA, Siqueira VLD, Scodro RBL, Cardoso RF, Caleffi-Ferracioli KR, 2016. Critical analysis: use of polymerase chain reaction to diagnose leprosy. Braz J Pharm Sci 52: 163–169.
-
30.
Reithinger R, Dujardin JC, 2007. Molecular diagnosis of leishmaniasis: current status and future applications. J Clin Microbiol 45: 21–25.
-
31.
Liu N, Zou D, DongD, Yang Z, Ao D, Liu W, Huang L, 2017. Development of a multiplex loop-mediated isothermal amplification method for the simultaneous detection of Salmonella spp. and Vibrio parahaemolyticus. Sci Rep 7: 45601.
-
32.
Aonuma H, Yoshimura A, Kobayashi T, Okado K, Badolo A, Nelson B, Kanukaa H, Fukumoto S, 2010. A single fluorescence-based LAMP reaction for identifying multiple parasites in mosquitoes. Exp Parasitol 25: 179–183.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 2274 | 826 | 33 |
| Full Text Views | 492 | 18 | 0 |
| PDF Downloads | 341 | 23 | 0 |
Rapid Multiplex Loop-Mediated Isothermal Amplification (m-LAMP) Assay for Differential Diagnosis of Leprosy and Post–Kala-Azar Dermal Leishmaniasis
Shweta Joshi
Shweta Joshi
Molecular Parasitology Laboratory, ICMR-National Institute of Pathology, Safdarjung Hospital Campus, New Delhi, India;
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Keerti K. Dixit
Keerti K. Dixit
Molecular Parasitology Laboratory, ICMR-National Institute of Pathology, Safdarjung Hospital Campus, New Delhi, India;
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Vanila Sharma
Vanila Sharma
Molecular Parasitology Laboratory, ICMR-National Institute of Pathology, Safdarjung Hospital Campus, New Delhi, India;
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V. Ramesh
V. Ramesh
Department of Dermatology, Safdarjung Hospital, New Delhi, India
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Ruchi Singh
Ruchi Singh
Molecular Parasitology Laboratory, ICMR-National Institute of Pathology, Safdarjung Hospital Campus, New Delhi, India;
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Poonam Salotra
Poonam Salotra
Molecular Parasitology Laboratory, ICMR-National Institute of Pathology, Safdarjung Hospital Campus, New Delhi, India;
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Abstract.
Leprosy and post–kala-azar dermal leishmaniasis (PKDL) are co-endemic neglected tropical diseases often misdiagnosed because of close resemblance in their clinical manifestations. The test that aids in differential diagnosis of leprosy and PKDL would be useful in endemic areas. Here, we report development of a multiplex loop-mediated isothermal amplification (m-LAMP) assay for differential detection of Mycobacterium leprae and Leishmania donovani using a real-time fluorometer. The m-LAMP assay was rapid with a mean amplification time of 15 minutes, and analytical sensitivity of 1 fg for L. donovani and 100 fg for M. leprae. The distinct mean Tm values for M. leprae and L. donovani allowed differentiation of the two organisms in the m-LAMP assay. Diagnostic sensitivity of the assay was evaluated by using confirmed cases of leprosy (n = 40) and PKDL (n = 40) (tissue and slit aspirate samples). All the leprosy and PKDL samples used in this study were positive by organism-specific QPCR and loop-mediated isothermal amplification assays. The diagnostic sensitivity of the m-LAMP assay was 100% (95% CI: 91.2–100.0%) for detecting PKDL and 95% for leprosy (95% CI: 83.1–99.4%). Our m-LAMP assay was successfully used to detect both M. leprae and L. donovani in a patient coinfected with leprosy and macular PKDL. The m-LAMP assay is rapid, accurate, and applicable for differential diagnosis of leprosy versus PKDL, especially in endemic areas.
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Author Notes
Financial suppot: The research was funded by the Indian Council of Medical Research (ICMR), New Delhi, India (intramural project). S. J. received grant from the Science and Engineering Research Board (SERB), Department of Science and Technology, Government of India, in the form of SERB-National Post-Doctoral Fellowship (Grant no. PDF/2016/003200).
Authors’ addresses: Shweta Joshi, Keerti K. Dixit, Vanila Sharma, Ruchi Singh, and Poonam Salotra, National Institute of Pathology, Molecular Parasitology Laboratory, New Delhi, India, E-mails: shweta26jan83@gmail.com, keerti.dixit@gmail.com, vanilasharma81@gmail.com, ruchisp@gmail.com, and poonamsalotra@hotmail.com. V. Ramesh, Dermatology and Venereology, Vardhman Mahavir Medical College and Safdarjung Hospital, New Delhi, India, E-mail: weramesh@gmail.com.
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1.
Lastória JC, Morgado de Abreu MAM, 2014. Leprosy: review of the epidemiological, clinical, and etiopathogenic aspects—part 1. An Bras Dermatol 89: 205–218.
-
2.
Rao PN, Suneetha S, 2018. Current situation of leprosy in India and its future implications. Indian Dermatol Online J 9: 83–89.
-
3.
National Leprosy Eradication Programme, 2018. NLEP Annual data (2017–18). Available at: http://nlep.nic.in. Accessed February 22, 2019.
-
4.
World Health Organisation, 2019. Leishmaniasis Facts Sheet. Available at: https://www.who.int/news-room/fact-sheets/detail/leishmaniasis. Accessed April 12, 2019.
-
5.
Mukherjee A, Ramesh V, Mishra RS, 1993. Post kala-azar dermal leishmaniasis: a light and electron microscopic study of 18 cases. J Cutan Pathol 20: 320–325.
-
6.
World Health Organization, 2010. Control of the leishmaniases, World Health Organ Tech Rep Ser 949: 22–26.
-
7.
Trindade MA, Silva LL, Braz LM, Amato VS, Naafs B, Sotto MN, 2015. Post-kala-azar dermal leishmaniasis and leprosy: case report and literature review. BMC Infect Dis 15: 543.
-
8.
Ramesh V, Ramam M, Singh R, Salotra P, 2008.Hypopigmented post-kala-azar dermal leishmaniasis. Int J Dermatol 47: 414–416.
-
9.
Khandpur S, Ramam M, Sharma VK, Salotra P, Singh MK, Malhotra A, 2003. Nerve involvement in Indian post kala-azar dermal leishmaniasis. Acta Derm Venereol 84: 245–246.
-
10.
Verma S, Kumar R, Katara GK, Singh LC, Negi NS, Ramesh V, Salotra P, 2010. Quantification of parasite load in clinical samples of leishmaniasis patients: IL-10 level correlates with parasite load in visceral leishmaniasis. PLoS One 5: e10107.
-
11.
Ghosh P et al. 2018. Evaluation of real-time PCR for diagnosis of post-kala-azar dermal leishmaniasis in endemic Foci of Bangladesh. Open Forum Infect Dis 5: ofy234.
-
12.
Bhargava A, Ramesh V, Verma S, Salotra P, Bala M, 2018. Revisiting the role of the slit-skin smear in the diagnosis of Indian post-kala-azar dermal leishmaniasis. Indian J Dermatol Venereol Leprol 84: 690–695.
-
13.
Azevedo MC, Ramuno NM, Fachin LR, Tassa M, Rosa PS, Belone AF, Diório SM, Soares CT, Garlet GP, Trombone AP, 2017. QPCR detection of Mycobacterium leprae in biopsies and slit skin smear of different leprosy clinical forms. Braz J Infect Dis 21: 71–78.
-
14.
Torres P, Camarena JJ, Gomez JR, Nogueira JM, Gimeno V, Navarro JC, Olmos A, 2003. Comparison of PCR mediated amplification of DNA and the classical methods for detection of Mycobacterium leprae in different types of clinical samples in leprosy patients and contacts. Lepr Rev 74: 18–30.
-
15.
Cheng X et al. 2019. Development and evaluation of a droplet digital PCR assay for the diagnosis of paucibacillary leprosy in skin biopsy specimens. PLoS Negl Trop Dis 13: e0007284.
-
16.
Barbieri RR et al. 2019. Quantitative polymerase chain reaction in paucibacillary leprosy diagnosis: a follow-up study. PLoS Negl Trop Dis 13: e0007147.
-
17.
De Paz HD, Brotons P, Muñoz-Almagro C, 2014. Molecular isothermal techniques for combating infectious diseases: towards low-cost point-of-care diagnostics. Expert Rev Mol Diagn 14: 827–843.
-
18.
Bühlmann A, Pothier JF, Rezzonico F, Smits TH, Andreou M, Boonham N, Duffy B, Frey JE, 2013. Erwiniaamylovora loop-mediated isothermal amplification (LAMP) assay for rapid pathogen detection and on-site diagnosis of fire blight. J Microbiol Methods 92: 332–339.
-
19.
Tomlinson JA, Dickinson MJ, Boonham N, 2010. Detection of Botrytis cinerea by loop-mediated isothermal amplification. Lett Appl Microbiol 51: 650–657.
-
20.
Lucchi NW, Demas A, Narayanan J, Sumari D, Kabanywanyi A, Kachur SP, Barnwell JW, Udhayakumar V, 2010. Real-time fluorescence loop mediated isothermal amplification for the diagnosis of malaria. PLoS One 29: e13733.
-
21.
Chaitanya VS, Cuello L, Das M, Sudharsan A, Ganesan P, Kanmani K, Rajan L, Ebenezer M, 2017. Analysis of a novel multiplex polymerase chain reaction assay as a sensitive tool for the diagnosis of indeterminate and tuberculoid forms of leprosy. Int J Mycobacteriol 6: 1–8.
-
22.
Verma S, Singh R, Sharma V, Bumb RA, Negi NS, Ramesh V, Salotra P, 2017. Development of a rapid loop-mediated isothermal amplification assay for diagnosis and assessment of cure of Leishmania infection. BMC Infect Dis 17: 223.
-
23.
Verma S, Avishek K, Sharma V, Negi NS, Ramesh V, Salotra P, 2013. Application of loop-mediated isothermal amplification assay for the sensitive and rapid diagnosis of visceral leishmaniasis and post-kala-azar dermal leishmaniasis. Diagn Microbiol Infect Dis 75: 390–395.
-
24.
Truman RW, Andrews PK, Robbins NY, Adams LB, Krahenbuhl JL, Gillis TP, 2008. Enumeration of Mycobacterium leprae using real-time PCR. PLoS Negl Trop Dis 2: e328.
-
25.
Singh A, Ramesh V, 2013. Histopathological features in leprosy, post-kala-azar dermal leishmaniasis, and cutaneous leishmaniasis. Indian J Dermatol Venereol Leprol 79: 360–366.
-
26.
Shao Y, Zhu S, Jin C, Chen F, 2011. Development of multiplex loop-mediated isothermal amplification-RFLP (mLAMP-RFLP) to detect Salmonella spp. and Shigella spp. in milk. Int J Food Microbiol 148: 75–79.
-
27.
Jung JH et al. 2015. Combination of multiplex reverse-transcription loop-mediated isothermal amplification with an immunochromatographic strip for subtyping influenza A virus. Anal Chim Acta 853: 541–547.
-
28.
Ball CS, Light YK, Koh CY, Wheeler SS, Coffey LL, Meagher RJ, 2016. Quenching of unincorporated amplification signal reporters in reverse-transcription loop-mediated isothermal amplification enabling bright, single-step, closed-tube, and multiplexed detection of RNA viruses. Anal Chem 88: 3562–3568.
-
29.
Maltempe FG, Baldin VP, Lopes MA, Siqueira VLD, Scodro RBL, Cardoso RF, Caleffi-Ferracioli KR, 2016. Critical analysis: use of polymerase chain reaction to diagnose leprosy. Braz J Pharm Sci 52: 163–169.
-
30.
Reithinger R, Dujardin JC, 2007. Molecular diagnosis of leishmaniasis: current status and future applications. J Clin Microbiol 45: 21–25.
-
31.
Liu N, Zou D, DongD, Yang Z, Ao D, Liu W, Huang L, 2017. Development of a multiplex loop-mediated isothermal amplification method for the simultaneous detection of Salmonella spp. and Vibrio parahaemolyticus. Sci Rep 7: 45601.
-
32.
Aonuma H, Yoshimura A, Kobayashi T, Okado K, Badolo A, Nelson B, Kanukaa H, Fukumoto S, 2010. A single fluorescence-based LAMP reaction for identifying multiple parasites in mosquitoes. Exp Parasitol 25: 179–183.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 2274 | 826 | 33 |
| Full Text Views | 492 | 18 | 0 |
| PDF Downloads | 341 | 23 | 0 |