Abstract
Leishmania infantum is the most common cause of visceral leishmaniasis (VL) in Iran, where mainly the patients are children under the age of 5 years. Timely, less invasive, and accurate diagnosis and proper treatment of the disease are necessary. This retrospective study aimed to search for a less invasive but robust algorithm on VL diagnostic tests in children. Four hundred and fifteen patients with clinical suspicion of VL, 50 healthy children from VL endemic areas, 46 healthy individuals from non-endemic VL areas, and 47 non-VL diseases were tested using three diagnostic tests: indirect immunofluorescent antibody test (IFAT), rK39-rapid diagnostic test (rK39-RDT), and quantitative PCR (qPCR). One hundred and two suspected VL cases were positive in at least one test and were cured after receiving appropriate treatment. Of these 102 VL patients, 94 were positive in qPCR, 84 in IFAT, and 79 in rK39-RDT. None of the tests detected all the patients, but overall, qPCR is capable of detecting more VL patients than serological tests, i.e., 92.2%, compared to IFAT, 82.4%, and rK39, 77.5%. There was only a significant difference between the sensitivity of qPCR and rK39-RDT (p = 0.024). The specificity was 100% for qPCR and IFAT (≥128) and 98.6% for rK39-RDT. qPCR alone is capable of detecting most of the VL-suspected children. Serological tests like IFAT and rk39-RDT are recommended to increase the overall sensitivity of detection in patients with a negative molecular test. Combining qPCR with a serological test (IFAT or rK39-RDT) can help diagnose 98% of VL. In laboratories without molecular facilities, we recommend testing with the combination of rK39-RDT and IFAT yielding a combined sensitivity of 93.1% equivalent to that of qPCR in our study.
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References
Alborzi A, Rasouli M, Nademi Z, Kadivar M, Pourabbas B (2006) Evaluation of rK39 strip test for the diagnosis of visceral leishmaniasis in infants. Eastern Mediterranean Health Journal 12(3-4):294–299
Alborzi A, Pourabbas B, Shahian F, Mardaneh J, Pouladfar GR, Ziyaeyan M (2008) Detection of Leishmania infantum kinetoplast DNA in the whole blood of asymptomatic individuals by PCR-ELISA and comparison with other infection markers in endemic areas, southern Iran. Am J Trop Med Hyg 79:839–842
Alvar J et al (2012) Leishmaniasis worldwide and global estimates of its incidence PloS one 7:e35671
Ashford DA et al (1995) Comparison of the polymerase chain reaction and serology for the detection of canine visceral leishmaniasis. Am J Trop Med Hyg 53:251–255
Bhattacharyya T, Boelaert M, Miles MA (2013) Comparison of visceral leishmaniasis diagnostic antigens in African and Asian Leishmania donovani reveals extensive diversity and region-specific polymorphisms. PLoS Negl Trop Dis 7:e2057
Boelaert M, Criel B, Leeuwenburg J, Van Damme W, Le Ray D, Van Der Stuyft P (2000) Visceral leishmaniasis control: a public health perspective. Trans R Soc Trop Med Hyg. 94:465–471
Boelaert M, el-Safi S, Hailu A, Mukhtar M, Rijal S, Sundar S, Wasunna M, Aseffa A, Mbui J, Menten J, Desjeux P, Peeling RW (2008) Diagnostic tests for kala-azar: a multi-centre study of the freeze-dried DAT, rK39 strip test and KAtex in East Africa and the Indian subcontinent. Trans R Soc Trop Med Hyg. 102:32–40
Chappuis F, Mueller Y, Nguimfack A, Rwakimari JB, Couffignal S, Boelaert M, Cavailler P, Loutan L, Piola P (2004) Diagnostic accuracy of two rK39 antigen-based dipsticks and the formol gel test for rapid diagnosis of visceral leishmaniasis in northeastern Uganda. J Clin Microbiol 43(12):5973–5977
Chappuis F, Rijal S, Soto A, Menten J, Boelaert M (2006) A meta-analysis of the diagnostic performance of the direct agglutination test and rK39 dipstick for visceral leishmaniasis. BMJ 333:723
Chappuis F, Sundar S, Hailu A, Ghalib H, Rijal S, Peeling RW, Alvar J, Boelaert M (2007) Visceral leishmaniasis: what are the needs for diagnosis, treatment and control? Nat. Rev. Microbiol. 5:S7–S16
Cunningham J, Hasker E, Das P, el Safi S, Goto H, Mondal D, Mbuchi M, Mukhtar M, Rabello A, Rijal S, Sundar S, Wasunna M, Adams E, Menten J, Peeling R, Boelaert M, for the WHO/TDR Visceral Leishmaniasis Laboratory Network (2012) A global comparative evaluation of commercial immunochromatographic rapid diagnostic tests for visceral leishmaniasis. Clin. Infect. Dis. 55:1312–1319
de Paiva-Cavalcanti M, de Morais RCS, Pessoa-e-Silva R, Trajano-Silva LAM, Gonçalves-de-Albuquerque SC, Tavares DHC, Brelaz-de-Castro MCA, Silva RF, Pereira VRA (2015) Leishmaniases diagnosis: an update on the use of immunological and molecular tools. Cell Biosci. 5:31
De Ruiter C, Van der Veer C, Leeflang M, Deborggraeve S, Lucas C, Adams E (2014) Molecular tools for diagnosis of visceral leishmaniasis: systematic review and meta-analysis of diagnostic test accuracy. J. Clin. Microbiol. 52:3147–3155
Deininger P (1990) Molecular cloning: a laboratory manual: Edited by J. Sambrook, EF Fritsch, and T. Maniatis. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1989 (in 3 volumes). Academic Press,
Elmahallawy EK, Martínez AS, Rodriguez-Granger J, Hoyos-Mallecot Y, Agil A, Mari JMN, Fernández JG (2014) Diagnosis of leishmaniasis. J. Infect. Dev. Ctries. 8:961–972
Fraga DBM, Pacheco LV, Borja LS, Tuy PGSE, Bastos LA, Solcà MS, Amorim LDAF, Veras PST (2016) The rapid test based on Leishmania infantum chimeric rK28 protein improves the diagnosis of canine visceral leishmaniasis by reducing the detection of false-positive dogs. PLoS Negl Trop Dis 10:e0004333
Freire ML, Machado de Assis T, Oliveira E, Moreira de Avelar D, Siqueira IC, Barral A, Rabello A, Cota G (2019) Performance of serological tests available in Brazil for the diagnosis of human visceral leishmaniasis. PLoS Negl Trop Dis 7:e0007484
Ghatee MA, Mirhendi H, Karamian M, Taylor WR, Sharifi I, Hosseinzadeh M, Kanannejad Z (2018) Population structures of Leishmania infantum and Leishmania tropica the causative agents of kala-azar in Southwest Iran. Parasitol. Res. 117:3447–3458
Junior MSCL et al (2013) Sensitivity of PCR and real-time PCR for the diagnosis of human visceral leishmaniasis using peripheral blood. Asian Pac. J. Trop. Dis. 3:10–15
Maia Z, Lírio M, Mistro S, Mendes CMC, Mehta SR, Badaro R (2012) Comparative study of rK39 Leishmania antigen for serodiagnosis of visceral leishmaniasis: systematic review with meta-analysis. PLoS Negl Trop Dis. 6:e1484
Mary C, Faraut F, Lascombe L, Dumon H (2004) Quantification of Leishmania infantum DNA by a real-time PCR assay with high sensitivity. J. Clin. Microbiol. 42:5249–5255
Mathis A, Deplazes P (1995) PCR and in vitro cultivation for detection of Leishmania spp. in diagnostic samples from humans and dogs. J. Clin. Microbiol. 33:1145–1149
Mengistu G, Kiessling R, Akuffo H (1990) The value of a direct agglutination test in the diagnosis of cutaneous and visceral leishmaniasis in Ethiopia. Trans R Soc Trop Med Hyg. 84:359–362
Mohebali M (2013) Visceral leishmaniasis in Iran: review of the epidemiological and clinical features. Iran J Parasitol. 8:348–358
Mohebali M, Edrissian GH, Shirzadi MR, Akhoundi B, Hajjaran H, Zarei Z et al (2011) An observational study on the current distribution of visceral leishmaniasis in different geographical zones of Iran and implication to health policy. Travel Med Infect Dis 9:67–74
Petrela R, Kuneshka L, Foto E, Zavalani F, Gradoni L (2010) Pediatric visceral leishmaniasis in Albania: a retrospective analysis of 1,210 consecutive hospitalized patients (1995–2009) PLoS Negl Trop Dis 4:e814
Pourabbas B, Moghadam AG, Pouladfar G, Rezaee Z, Alborzi A (2013) Quantification of Leishmania infantum kinetoplast DNA for monitoring the response to meglumine antimoniate therapy in visceral leishmaniasis. Am J Trop Med Hyg 88:868–871
Rezaei Z et al (2019) Expression of a rK39 homologue from an Iranian Leishmania infantum isolate in Leishmania tarentolae for serodiagnosis of visceral leishmaniasis. Parasites Vectors 12:1–9
Sarkari B, Naraki T, Ghatee MA, Abdolahi Khabisi S, Davami MH (2016) Visceral leishmaniasis in Southwestern Iran: a retrospective clinico-hematological analysis of 380 consecutive hospitalized cases (1999–2014). PloS one 11:e0150406
Sarkari B, Rezaei Z, Mohebali M (2018) Immunodiagnosis of visceral leishmaniasis: current status and challenges: a review article. Iran J Parasitol. 13:331
Savoia D (2015) Recent updates and perspectives on leishmaniasis. J. Infect. Dev. Ctries. 9:588–596
Simon AK, Hollander GA, McMichael A (2015) Evolution of the immune system in humans from infancy to old age. Proc. Royal Soc. B 282:20143085
Srividya G, Kulshrestha A, Singh R, Salotra P (2012) Diagnosis of visceral leishmaniasis: developments over the last decade. Parasitol. Res. 110:1065–1078
Sudarshan M, Sundar S (2014) Parasite load estimation by qPCR differentiates between asymptomatic and symptomatic infection in Indian visceral leishmaniasis. Diagn. Microbiol. Infect. Dis. 80:40–42
Sundar S, More DK, Singh MK, Singh VP, Sharma S, Makharia A, Kumar PCK, Murray HW (2000) Failure of pentavalent antimony in visceral leishmaniasis in India: report from the center of the Indian epidemic. Clin. Infect. Dis. 31:1104–1107
Sundar S, Singh RK, Bimal SK, Gidwani K, Mishra A, Maurya R, Singh SK, Manandhar KD, Boelaert M, Rai M (2007) Comparative evaluation of parasitology and serological tests in the diagnosis of visceral leishmaniasis in India: a phase III diagnostic accuracy study. Trop. Med. Int. Health. 12:284–289
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Our thanks are due to Hassan Khajehei for copy editing of the manuscript.
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The study was financially supported by the Professor Alborzi Clinical Microbiology Research Center, Shiraz University of Medical Sciences. VK received funding from the European Union’s Horizon 2020 research and innovation program under the Marie Sklodowska-Curie grant agreement No. 642609. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
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Rezaei, Z., Pourabbas, B., Asaei, S. et al. Pediatric visceral leishmaniasis: a retrospective study to propose the diagnostic tests algorithm in southern Iran. Parasitol Res 120, 1447–1453 (2021). https://doi.org/10.1007/s00436-021-07067-1
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DOI: https://doi.org/10.1007/s00436-021-07067-1