Mediterranean visceral leishmaniasis : update on biological diagnosis
##plugins.themes.academic_pro.article.main##
Abstract
Visceral leishmaniasis (VL) is a severe life threatening parasitosis requiring early management of cases. It is an emerging disease in the Mediterranean region with a spread of endemic areas and an increase in case incidence. The patient profile has also evolved with more affected adults, presenting generally non-specific symptoms. Hence the interest of a systematic biological confirmation.
The microscopic detection of Leishmania amastigotes in bone marrow aspirates (BMA) smears is the gold standard diagnostic technique. However, it requires invasive sampling. Serological tests searching for specific antibodies remain highly contributory, but their interpretation must always take into account the epidemiological context and the patient’s clinical and biological features. Currently, the Western-Blot represents the most specific serological technique for diagnostic confirmation. VL diagnosis has greatly improved by the introduction of both rapid diagnostic tests (RDTs) and molecular biological techniques. RDTs using recombinant rk39 antigen are easy to perform and deliver results in less than 30 minutes. Real-time PCR (Polymerase Chain Reaction) is currently retained as the best technique for VL diagnosis. It is efficient on simple blood samples, allowing to avoid invasive BMA needed for microscopy. In addition, real time PCR estimates parasite load which is helpful for the post-treatment follow-up. In any case, the choice of techniques to be used should be strategic and adapted to the local epidemiology as well as to the means available.
##plugins.themes.academic_pro.article.details##
References
- Burza S, Croft SL, Boelaert M. Leishmaniasis. Lancet Lond Engl 2018;392(10151):951-70.
- Alvar J, Aparicio P, Aseffa A, Den Boer M, Cañavate C, Dedet J-P, et al. The relationship between leishmaniasis and AIDS: the second 10 years. Clin Microbiol Rev 2008;21(2):334-59.
- Antinori S, Cascio A, Parravicini C, Bianchi R, Corbellino M. Leishmaniasis among organ transplant recipients. Lancet Infect Dis 2008;8(3):191-9.
- Cruz I, Morales M, Noguer I, Rodríguez-Arenas MÁ, Alvar J. Leishmania in discarded syringes from intravenous drug users. Lancet 2002;359:1124-5.
- Pagliano P, Carannante N, Rossi M, Gramiccia M, Gradoni L, Faella FS, et al. Visceral leishmaniasis in pregnancy: a case series and a systematic review of the literature. J Antimicrob Chemother 2005;55(2):229-33.
- Savoia D. Recent updates and perspectives on leishmaniasis. J Infect Dev Ctries 2015;9(6):588-96.
- World Health Organization. Control of the leishmaniases. World Health Organ Tech Rep Ser. 2010;(949):xii‐xiii, 1-186.
- Gangneux J-P, Belaz S, Robert-Gangneux F. Mise au point et actualités sur la leishmaniose viscérale méditerranéenne.J Anti-Infect 2015;17(1):25‐8.
- Faucher B, Piarroux R. Visceral leishmaniasis: an update.Rev Med Interne 2011;32(9):544‐51.
- Aoun K, Jeddi F, Amri F, Ghrab J, Bouratbine A. Current epidemiological data on visceral leishmaniasis in Tunisia. Med Mal Infect 2009;39(10):775-9.
- Ben Salah AB, Ben Ismail R, Amri F, Chlif S, Ben Rzig F, Kharrat H, et al. Investigation of the spread of human visceral leishmaniasis in central Tunisia. Trans R Soc Trop Med Hyg 2000;94(4):382-6.
- Benabid M, Ghrab J, Rhim A, Ben-Romdhane R, Aoun K, Bouratbine A. Temporal dynamics and Leishmania infantum infection prevalence of Phlebotomus perniciosus (Diptera, Phlebotominae) in highly endemic areas of visceral leishmaniasis in Tunisia. PloS One 2017;12(9):e0184700.
- Adel A, Boughoufalah A, Saegerman C, De Deken R, Bouchene Z, Soukehal A, et al. Epidemiology of visceral leishmaniasis in Algeria: an update. PloS One 2014;9(6):e99207.
- Kahime K, Boussaa S, Nhammi H, Boumezzough A. Urbanization of human visceral leishmaniasis in Morocco. Parasite Epidemiol Control 2017;2(4):1-6.
- Kholoud K, Bounoua L, Sereno D, El Hidan M, Messouli M. Emerging and Re-Emerging Leishmaniases in the Mediterranean Area: What Can Be Learned from a Retrospective Review Analysis of the Situation in Morocco during 1990 to 2010? Microorganisms 2020;8(10):E1511.
- Mniouil M, Fellah H, Amarir F, Et-Touys A, Bekhti K, Adlaoui EB, et al. Epidemiological characteristics of visceral leishmaniasis in Morocco (1990-2014): an update. Acta Trop 2017;170:169-77.
- Bouratbine A, Aoun K, Chahed MK, Ben Ismail R. Données épidémiologiques sur la leishmaniose viscérale infantile en Tunisie en 1993. Med Mal Infect 1998;28(5):446‐7.
- Harrat Z, Addadi K, Belkaid M, Tabet-Derraz O. Visceral leishmaniasis in Algeria. Cases reported of visceral leishmaniasis (1985-1990). Bull Soc Pathol Exot 1990 1992;85(4):296-301.
- Marty P. Visceral leishmaniases: epidemiology and diagnosis. Med Mal Infect 2005;35 Suppl 2:S72-73.
- Essabbah Aguir N, Toumi A, Loussaïf C, Gorcii M, M’rad S, Ben Brahim H et al. La leishmaniose viscérale de l’adulte immunocompétent : à propos de six cas. Pathologie Biologie.2013;61 (2):54-58.
- Aoun K, Kooli C, Bouratbine A, Ben Romdhane N, Kaaroud H, Ben Maı̈ z H, et al. Aspects épidémiologiques et cliniques de la leishmaniose viscérale de l’adulte en Tunisie. Med Mal Infect 2002;32(7):387-92.
- Aoun K, Kaaroud H, Hamzaoui S, Siala E, Kooli C, Turki S, et al. Particularities of visceral leishmaniasis in adults not infected by HIV in Tunisia. Med Trop 2004;64(2):160-2.
- Dedet J-P, Carme B, Desbois N, Bourdoiseau G, Lachaud L, Pratlong F. Épidémiologie des leishmanioses autochtones en France métropolitaine et d’outre-mer. Presse Med 2013;42(11):1469-81.
- Marty P, Delaunay P, Fissore C, Le Fichoux Y. Mediterranean leishmaniasis caused by Leishmania infantum. Update on the utility of the IT-Leish and ID-Pagia leishmaniasis tests. Med Trop 2007;67(1):79-85.
- Lachaud L, Dedet JP, Marty P, Faraut F, Buffet P, Gangneux JP, et al. Surveillance of leishmaniases in France, 1999 to 2012. Euro Surveill 2013;18:20534.
- Alvar J, Vélez ID, Bern C, Herrero M, Desjeux P, Cano J, et al. Leishmaniasis worldwide and global estimates of its incidence. PloS One 2012;7(5):e35671.
- Abdalmaula GH, Barbadoro P, Marigliano A, Illuminati D, Di Stanislao F, D’Errico MM, et al. Human visceral leishmaniasis: a picture from Italy. J Infect Public Health 2013;6(6):465-72.
- Bourdoiseau G. Le vétérinaire acteur de santé publique : exemple de la leishmaniose canine, zoonose vectorielle. Bull Académie Natl Médecine 2015;199(6):909-20.
- Murray HW, Berman JD, Davies CR, Saravia NG. Advances in leishmaniasis. Lancet Lond Engl 2005;366(9496):1561-77.
- Riera C, Fisa R, López-Chejade P, Serra T, Girona E, Jiménez M, et al. Asymptomatic infection by Leishmania infantum in blood donors from the Balearic Islands (Spain). Transfusion 2008;48(7):1383-9.
- Kallel K, Ammari L, Kaouech E, Belhadj S, Anane S, Kilani B, et al. Portage asymptomatique de Leishmania infantum chez des malades tunisiens infectés par le VIH. Pathologie Biologie 2007;55(10):521-4.
- Louzir H, Dellagi K. Les leishmanioses: un modèle d’étude des interactions hôte-parasite; implications pour la maladie humaine. Ann Inst Pasteur Actual 1999;10:67-80.
- Desjeux P. Leishmaniasis: current situation and new perspectives. Comp Immunol Microbiol Infect Dis 2004;27(5):305-18.
- Pace D. Leishmaniasis. J Infect 2014;69 Suppl 1:S10-18.
- Zayet S, Abdelmalek R, Ben Romdhane N, Sahli H, Aoun K, Tiouiri Benaissa H. Isolated cervical lymphadenopathy: Do not forget atypical leishmaniasis. Presse Med 2017;46(4):452-4.
- Zougaghi L, Moutaj R, Chabaa L, Agoumi A. Infantile visceral Leishmaniasis: epidemiological, clinical and biological characteristics. About 93 case reports in the children hospital of Rabat. Arch Pediatr Organe Off Soc Francaise Pediatr 2009;16(11):1513-8.
- Aoun K, Kalboussi Y, Ben Sghaier I, Souissi O, Hammami H, Bellali H, et al. Assessment of incubation period of cutaneous leishmaniasis due to Leishmania major in Tunisia. Am J Trop Med Hyg 2020;103(5):1934-7.
- Deniau M, Cañavate C, Faraut-Gambarelli F, Marty P. The biological diagnosis of leishmaniasis in HIV-infected patients. Ann Trop Med Parasitol 2003;97 Suppl 1:115-33.
- Aissi W, Ben Hellel K, Habboul Z, Ben Sghaier I, Harrat Z, Bouratbine A, et al. Profils épidémiologique, clinique et biologique de la leishmaniose viscérale infantile à l’hôpital de Kairouan (Tunisie) : à propos de 240 cas. Bull Soc Pathol Exot 2015;108(4):265-71.
- Rosenthal E, Marty P, del Giudice P, Pradier C, Ceppi C, Gastaut JA, et al. HIV and Leishmania coinfection: a review of 91 cases with focus on atypical locations of Leishmania. Clin Infect Dis 2000;31:1093-5.
- Zijlstra EE, Ali MS, el-Hassan AM, el-Toum IA, Satti M, Ghalib HW, et al. Kala-azar: a comparative study of parasitological methods and the direct agglutination test in diagnosis. Trans R Soc Trop Med Hyg 1992;86(5):505‑7.
- Srivastava P, Dayama A, Mehrotra S, Sundar S. Diagnosis of visceral leishmaniasis. Trans R Soc Trop Med Hyg 2011;105(1):1-6.
- Chappuis F, Sundar S, Hailu A, Ghalib H, Rijal S, Peeling RW, et al. Visceral leishmaniasis: what are the needs for diagnosis, treatment and control? Nat Rev Microbiol 2007;5(11):873-82.
- Sakkas H, Gartzonika C, Levidiotou S. Laboratory diagnosis of human visceral leishmaniasis. J Vector Borne Dis 2016;53(1):8-16.
- Sarkari B, Rezaei Z, Mohebali M. Immunodiagnosis of Visceral Leishmaniasis: Current Status and Challenges: A Review Article. Iran J Parasitol 2018;13(3):331-41.
- Srividya G, Kulshrestha A, Singh R, Salotra P. Diagnosis of visceral leishmaniasis: developments over the last decade. Parasitol Res 2012;110(3):1065-78.
- Elmahallawy EK, Sampedro Martinez A, Rodriguez-Granger J, Hoyos-Mallecot Y, Agil A, Navarro Mari JM, et al. Diagnosis of leishmaniasis. J Infect Dev Ctries 2014;8(8):961-72.
- Izri MA, Deniau M, Brière C, Rivollet D, Petithory JC, Houin R, et al. Leishmaniasis in AIDS patients: results of leukocytoconcentration, a fast biological method of diagnosis. Bull World Health Organ 1996;74(1):91-3.
- Diro E, Yansouni CP, Takele Y, Mengesha B, Lynen L, Hailu A, et al. Diagnosis of visceral leishmaniasis using peripheral blood microscopy in Ethiopia: A Prospective P h a s e - I I I study of the diagnostic performance of different concentration techniques compared to tissue aspiration. Am J Trop Med Hyg 2017;96(1):190-6.
- Chemli J, Abroug M, Fathallah A, Abroug S, Ben Said M, Harbi A. Contribution of leukoconcentration in the diagnosis of Kala-azar in Tunisia. Med Mal Infect 2006;36(7):390-2.
- Chouihi E, Amri F, Bouslimi N, Siala E, Selmi K, Zallagua N, et al. Cultures on NNN medium for the diagnosis of leishmaniasis. Pathol Biol 2009;57(3):219-24.
- Belhadj S, el Houda Toumi N, Kallel K, Dakhlia H, Pratlong F, Boussen N, et al. Visceral leishmaniasis in immunocompetent children: diagnostic and epidemiologic value of peripheral Leishmania blood culture. Tunis Med 2001;79(4):231-3.
- Aoun K, Bouratbine A, Harrat Z, Belkaïd M, Bel Hadj Ali S. Particular profile of the zymodemes of Leishmania infantum causing visceral leishmaniasis in Tunisia. Bull Soc Pathol Exot 2001;94(5):375-7.
- Rioux JA, Lanotte G, Serres E, Pratlong F, Bastien P, Perieres J. Taxonomy of Leishmania. Use of isoenzymes. Suggestions for a new classification. Ann Parasitol Hum Comp 1990;65(3):111-25.
- Carrió J, Portús M. In vitro susceptibility to pentavalent antimony in Leishmania infantum strains is not modified during in vitro or in vivo passages but is modified after host treatment with meglumine antimoniate. BMC Pharmacol 2002;2:11.
- Belhadj S, Hicheri-Helali J, Kallel K, Kaouech E, Abaza H, Toumi NEH, et al. Place de la culture dans le diagnostic parasitologique des leishmanioses viscérales et cutanées: Expérience tunisienne. Rev Fr Lab 2005;369:41-5.
- Belhadj S, Toumi NH, Dakhlia H, Kallel K, Boussen N, Ben Chaabane T, et al. Peripheral blood culture as a diagnostic modality for visceral leishmaniasis: apropos of 61 cases. Med Trop 2002;62(2):155-7.
- Sundar S, Rai M. Laboratory diagnosis of visceral leishmaniasis. Clin Diagn Lab Immunol 2002;9(5):951-8.
- Boelaert M, Bhattacharya S, Chappuis F, El Safi SH, Hailu A, Mondal D, et al. Evaluation of rapid diagnostic tests: visceral leishmaniasis. Nature Reviews Microbiology 2007;5(11):S31-9.
- Badaró R, Reed SG, Barral A, Orge G, Jones TC. Evaluation of the micro enzyme-linked immunosorbent assay (ELISA) for antibodies in American visceral leishmaniasis: antigen selection for detection of infection-specific responses. Am J Trop Med Hyg 1986;35(1):72-8.
- Kumar R, Pai K, Pathak K, Sundar S. Enzyme-linked immunosorbent assay for recombinant K39 antigen in diagnosis and prognosis of Indian visceral leishmaniasis. Clin Diagn Lab Immunol 2001;8(6):1220-4.
- Maia Z, Lírio M, Mistro S, Mendes CMC, Mehta SR, Badaro R. Comparative study of rK39 Leishmania antigen for serodiagnosis of visceral leishmaniasis: systematic review with meta-analysis. PLoS Negl Trop Dis 2012;6(1):e1484.
- Salotra P, Sreenivas G, Nasim AA, Subba Raju BV, Ramesh V. Evaluation of Enzyme-Linked Immunosorbent Assay for Diagnosis of Post-Kala-Azar Dermal Leishmaniasis with Crude or Recombinant k39 Antigen. Clin Diagn Lab Immunol 2002;9(2):370-3.
- Vaish M, Bhatia A, Reed SG, Chakravarty J, Sundar S. Evaluation of rK28 antigen for serodiagnosis of visceral Leishmaniasis in India. Clin Microbiol Infect 2012;18(1):81-5.
- Atta AM, D’Oliveira null, Correa J, Atta ML, Almeida RP, Carvalho EM. Anti-leishmanial IgE antibodies: a marker of active disease in visceral leishmaniasis. Am J Trop Med Hyg 1998;59(3):426-30.
- Saghrouni F, Gaïed-Meksi S, Fathallah A, Amri F, Ach H, Guizani I, et al. Immunochromatographic rK39 strip test in the serodiagnosis of visceral leishmaniasis in Tunisia. Trans R Soc Trop Med Hyg 2009;103(12):1273-8.
- Lévêque MF, Battery E, Delaunay P, Lmimouni BE, Aoun K, L’Ollivier C, et al. Evaluation of six commercial kits for the serological diagnosis of Mediterranean visceral leishmaniasis. PLoS Negl Trop Dis 2020;14(3):e0008139.
- Seyyedtabaei Sj, Rostami A, Haghighi A, Mohebali M, Kazemi B, Fallahi S, et al. Detection of potentially diagnostic Leishmania Antigens with Western Blot analysis of sera from patients with cutaneous and visceral leishmaniases. Iran J Parasitol 2017;12(2):206-14.
- Marty P, Lelièvre A, Quaranta JF, Suffia I, Eulalio M, GariToussaint M, et al. Detection by Western blot of four antigens characterizing acute clinical leishmaniasis due to Leishmania infantum. Trans R Soc Trop Med Hyg 1995;89(6):690-1.
- Fissore C, Delaunay P, Ferrua B, Rosenthal E, Del Giudice P, Aufeuvre J-P, et al. Convenience of serum for visceral leishmaniasis diagnosis by PCR. J Clin Microbiol 2004;42(11):5332-3.
- Saghrouni F, Khammari I, Kaabia N, Bouguila J, Ben Abdeljelil J, Fathallah A, et al. Asymptomatic carriage of Leishmania in family members of patients with visceral leishmaniasis in Central Tunisia. Pathol Biol 2012;60(5):e55-58.
- Cota GF, de Sousa MR, Demarqui FN, Rabello A. The diagnostic accuracy of serologic and molecular methods for detecting visceral leishmaniasis in HIV infected patients: meta-analysis. PLoS Negl Trop Dis 2012;6(5):e1665.
- Mohebali M, Keshavarz H, Shirmohammad S, Akhoundi B, Borjian A, Hassanpour G, et al. The diagnostic accuracy of direct agglutination test for serodiagnosis of human visceral leishmaniasis: a systematic review with meta-analysis. BMC Infect Dis 2020;20(1):946.
- Chappuis F, Rijal S, Soto A, Menten J, Boelaert M. A meta-analysis of the diagnostic performance of the direct agglutination test and rK39 dipstick for visceral leishmaniasis. BMJ 2006;333(7571):723.
- Bangert M, Flores-Chávez MD, Llanes-Acevedo IP, Arcones C, Chicharro C, García E, et al. Validation of rK39 immunochromatographic test and direct agglutination test for the diagnosis of Mediterranean visceral leishmaniasis in Spain. PLoS Negl Trop Dis 2018;12(3):e0006277.
- Kohanteb J, Ardehali S. Cross-reaction of sera from patients with various infectious diseases with Leishmania infantum. Med Princ Pract 2005;14(2):79-82.
- Maia Z, Lírio M, Mistro S, Mendes CMC, Mehta SR, Badaro R. Comparative study of rK39 Leishmania antigen for serodiagnosis of visceral leishmaniasis: systematic review with meta-analysis. PLoS Negl Trop Dis 2012;6(1):e1484.
- Akhoundi B, Mohebali M, Babakhan L, Edrissian G-H, Eslami M-B, Keshavarz H, et al. Rapid detection of human Leishmania infantum infection: a comparative field study using the fast agglutination screening test and the direct agglutination test. Travel Med Infect Dis 2010;8(5):305-10.
- Mohapatra TM, Singh DP, Sen MR, Bharti K, Sundar S. Compararative evaluation of rK9, rK26 and rK39 antigens in the serodiagnosis of Indian visceral leishmaniasis. J Infect Dev Ctries 2010;4(2):114-7.
- Farahmand M, Khalaj V, Mohebali M, Khalili G, Naderi S, Ghaffarinejad P, et al. Comparison of recombinant A2-ELISA with rKE16 dipstick and direct agglutination tests for diagnosis of visceral leishmaniasis in dogs in Northwestern Iran. Rev Soc Bras Med Trop 2015;48(2):188-93.
- Zijlstra EE, Nur Y, Desjeux P, Khalil EA, El-Hassan AM, Groen J. Diagnosing visceral leishmaniasis with the recombinant K39 strip test: experience from the Sudan. Trop Med Int Health 2001;6(2):108-13.
- Bhattacharyya T, Boelaert M, Miles MA. Comparison of visceral leishmaniasis diagnostic antigens in African and Asian Leishmania donovani reveals extensive diversity and region-specific polymorphisms. PLoS Negl Trop Dis 2013;7(2):e2057.
- Abass E, Bollig N, Reinhard K, Camara B, Mansour D, Visekruna A, et al. rKLO8, a novel Leishmania donovani - derived recombinant immunodominant protein for sensitive detection of visceral leishmaniasis in Sudan. PLoS Negl Trop Dis 2013;7(7):e2322.
- Van Griensven J, Diro E. Visceral Leishmaniasis: Recent Advances in Diagnostics and Treatment Regimens. Infect Dis Clin North Am 2019;33(1):79-99.
- Cunningham J, Hasker E, Das P, El Safi S, Goto H, Mondal D, et al. A Global comparative evaluation of commercial Immunochromatographic Rapid Diagnostic Tests for visceral leishmaniasis. Clin Infect Dis 2012;55(10):1312-9.
- Boelaert M, Verdonck K, Menten J, Sunyoto T, van Griensven J, Chappuis F, et al. Rapid tests for the diagnosis of visceral leishmaniasis in patients with suspected disease. Cochrane Database Syst Rev 2014;(6):CD009135.
- Mukhtar M, Abdoun A, Ahmed AE, Ghalib H, Reed SG, Boelaert M, et al. Diagnostic accuracy of rK28-based immunochromatographic rapid diagnostic tests for visceral leishmaniasis: a prospective clinical cohort study in Sudan. Trans R Soc Trop Med Hyg 2015;109(9):594-600.
- De Paiva-Cavalcanti M, de Morais RCS, Pessoa-E-Silva R, TrajanoSilva LAM, Gonçalves-de-Albuquerque S da C, Tavares D de HC, et al. Leishmaniases diagnosis: an update on the use of immunological and molecular tools. Cell Biosci 2015;5:31.
- Cota GF, de Sousa MR, de Freitas Nogueira BM, Gomes LI, Oliveira E, Assis TSM, et al. Comparison of parasitological, serological, and molecular tests for visceral leishmaniasis in HIVinfected patients: a cross-sectional delayed-type study. Am J Trop Med Hyg 2013;89(3):570-7.
- Sundar S, Maurya R, Singh RK, Bharti K, Chakravarty J, Parekh A, et al. Rapid, non invasive diagnosis of visceral leishmaniasis in India: comparison of two immunochromatographic strip tests for detection of anti-K39 antibody. J Clin Microbiol 2006;44(1):251-3.
- Singh S, Kumari V, Singh N. Predicting kala-azar disease manifestations in asymptomatic patients with latent Leishmania donovani infection by detection of antibody against recombinant K39 antigen. Clin Diagn Lab Immunol 2002;9(3):568-72.
- Sinha PK, Bimal S, Pandey K, Singh SK, Ranjan A, Kumar N, et al. A community-based, comparative evaluation of direct agglutination and rK39 strip tests in the early detection of subclinical Leishmania donovani infection. Ann Trop Med Parasitol 2008;102(2):119-25.
- Schallig HDFH, Canto-Cavalheiro M, da Silva ES. Evaluation of the direct agglutination test and the rK39 dipstick test for the sero-diagnosis of visceral leishmaniasis. Mem Inst Oswaldo 2002;97(7):1015-8.
- Romero GAS, de la Glória Orge Orge M, de Farias Guerra MV, Paes MG, de Oliveira Macêdo V, de Carvalho EM. Antibody response in patients with cutaneous leishmaniasis infected by Leishmania (Viannia) braziliensis or Leishmania (Viannia) guyanensis in Brazil. Acta Trop 2005;93(1):49-56.
- Kohanteb J, Ardehali S. Cross-reaction of sera from patients with various infectious diseases with Leishmania infantum. Med Princ Pract 2005;14(2):79-82.
- De Ruiter CM, van der Veer C, Leeflang MMG, Deborggraeve S, Lucas C, Adams ER. Molecular tools for diagnosis of visceral leishmaniasis: systematic review and meta-analysis of diagnostic test accuracy. J Clin Microbiol 2014;52(9):3147- 55.
- Singh OP, Hasker E, Sacks D, Boelaert M, Sundar S. Asymptomatic Leishmania infection: a new challenge for Leishmania control. Clin Infect Dis 2014;58(10):1424-9.
- Gidwani K, Picado A, Ostyn B, Singh SP, Kumar R, Khanal B, et al. Persistence of Leishmania donovani antibodies in past visceral leishmaniasis cases in India. Clin Vaccine Immunol 2011;18(2):346-8.
- Hommel M, Sarkari B, Carney J, Chance ML. Katex for the diagnosis of human visceral leishmaniasis. Med Trop 2001;61(6):503-5.
- Ben-Abid M, Galaï Y, Habboul Z, Ben-Abdelaziz R, BenSghaier I, Aoun K, et al. Diagnosis of Mediterranean visceral leishmaniasis by detection of Leishmania -related antigen in urine and oral fluid samples. Acta Tropica 2017;167:71-2.
- Ghosh P, Bhaskar KRH, Hossain F, Khan MAA, Vallur AC, Duthie MS, et al. Evaluation of diagnostic performance of rK28 ELISA using urine for diagnosis of visceral leishmaniasis. Parasit Vectors 2016;9(1):383.
- Vallur AC, Tutterrow YL, Mohamath R, Pattabhi S, Hailu A, Abdoun AO, et al. Development and comparative evaluation of two antigen detection tests for Visceral Leishmaniasis. BMC Infect Dis 2015;15:384.
- Riera C, Fisa R, Udina M, Gállego M, Portus M. Detection of Leishmania infantum cryptic infection in asymptomatic blood donors living in an endemic area (Eivissa, Balearic Islands, Spain) by different diagnostic methods. Trans R Soc Trop Med Hyg 2004;98(2):102-10.
- . García-García JA, Martín-Sánchez J, Gállego M, RiveroRomán A, Camacho A, Riera C, et al. Use of noninvasive markers to detect Leishmania infection in asymptomatic human immunodeficiency virus-infected patients. J Clin Microbiol 2006;44(12):4455-8.
- Cruz I, Nieto J, Moreno J, Cañavate C, Desjeux P, Alvar J. Leishmania/HIV co-infections in the second decade. Indian J Med Res 2006;123(3):357-88.
- . Sundar S, Singh OP. Molecular Diagnosis of Visceral Leishmaniasis. Mol Diagn Ther 2018;22(4):443-57.
- Galluzzi L, Ceccarelli M, Diotallevi A, Menotta M, Magnani M. Real-time PCR applications for diagnosis of leishmaniasis. Parasit Vectors 2018;11(1):273.
- Kaouech E, Kallel K, Toumi NH, Belhadj S, Anane S, Babba H, et al. Pediatric visceral leishmaniasis diagnosis in Tunisia: Comparative study between optimized PCR assays and parasitological methods. Parasite 2008;15:143-50.
- Mary C, Faraut F, Lascombe L, Dumon H. Quantification of Leishmania infantum DNA by a real-time PCR assay with high sensitivity. J Clin Microbiol 2004;42(11):5249-55.
- Mary C, Faraut F, Drogoul M-P, Xeridat B, Schleinitz N, Cuisenier B, et al. Reference values for Leishmania infantum parasitemia in different clinical presentations: quantitative polymerase chain reaction for therapeutic monitoring and patient follow-up. Am J Trop Med Hyg 2006;75(5):858-63.
- Aoun K, Chouihi E, Amri F, Ben Alaya N, Raies A, Mary C, et al. Short report: Contribution of quantitative realtime polymerase chain reaction to follow-up of visceral leishmaniasis patients treated with meglumine antimoniate. Am J Trop Med Hyg 2009;81(6):1004-6.
- Sudarshan M, Sundar S. Parasite load estimation by qPCR differentiates between asymptomatic and symptomatic infection in Indian visceral leishmaniasis. Diagn Microbiol Infect Dis 2014;80(1):40-2.
- Chaouch M, Aoun K, Ben Othman S, Ben Abid M, Ben Sghaier I, Bouratbine A, et al. Development and assessment of Leishmania major and Leishmania tropica specific LoopMediated Isothermal Amplification Assays for the diagnosis of cutaneous leishmaniasis in Tunisia. Am J Trop Med Hyg 2019;101(1):101-7.
- Verma S, Avishek K, Sharma V, Negi NS, Ramesh V, Salotra P. 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 2013;75(4):390-5.
- Bousslimi N, Ben Abda I, Ben Mously R, Siala E, Harrat Z, Zallagua N, et al. Place de l’identification des leishmanies par la polymerase chain reaction – restriction fragment length polymerase dans l’étude de l’épidémiologie des leishmanioses cutanées en Tunisie. Pathol Biol 2014;62(1):30-3.
- Castelli G, Bruno F, Caputo V, Fiorella S, Sammarco I, Lupo T, et al. Genetic tools discriminate strains of Leishmania infantum isolated from humans and dogs in Sicily, Italy. PLoS Negl Trop Dis 2020;14(7):e0008465.
- Dereure J, El-Safi SH, Bucheton B, Boni M, Kheir MM, Davoust B, et al. Visceral leishmaniasis in eastern Sudan: parasite identification in humans and dogs; hostparasite relationships. Microbes Infect 2003;5(12):1103-8.