*Corresponding Author Address: Prof Dr. Mohammed Sh. Jebur, Institute of Medical Tech. Email: d_mohamed_1959@yahoo.com World Journal of Pharmaceutical Sciences ISSN (Print): 2321-3310; ISSN (Online): 2321-3086 Published by Atom and Cell Publishers © All Rights Reserved Available online at: http://www.wjpsonline.org/ Original Article Comparison of IFAT, rk39 and PCR for diagnosis of kala-azar in Iraqi children Basheer Abdullah Naseralla*, Maher Ali Al-Quraishi**, Mohammed Sh. Jebur* *Institute of Medical Technology and **College of Science / Babylon University, Babylon, Iraq Received: 05-12-2014 / Revised: 26-12-2014 / Accepted: 26-01-2015 ABSTRACT A comparative study was carried out through IFAT, rk39 and PCR diagnostic methods for detection of pediatric visceral leishmaniasis (Kala-azar) in Baghdad hospitals during the period from July 2013 to September 2014. Blood samples from 178 suspected VL patients were examined by IFAT, rk39 and PCR. Results of polymerase chain reaction which depending on highly repetitive sequence regions, reported that only 39 (21.9%) of samples were positive to visceral leishmaniasis and 139 (79.1%) were negative. So that these results were considered as standard test for IFAT and rk39 test, for that the validity of IFAT according to PCR technique detected the sensitivity of IFAT was (100%) and specificity (82%) with accuracy (89%). While the validity of rK39 test according to PCR technique, it was found that the sensitivity of rK39 (97.4%) and specificity (85.2%) with accuracy (90%).The study concludes that these diagnostic methods had an excellent ability of discrimination in diagnosis of VL cases. Key words: Visceral leishmaniasis, IFAT, rk39 and PCR INTRODUCTION Leishmaniasis is endemic in 98 countries and 3 territories on five continents; it is estimated that there are 2 million new cases annually worldwide and up to 350 million people at risk of the disease [1,2]. The parasite invades internal organs such as spleen, liver and bone marrow which usually with mortality rate almost 100% if left untreated[3]. The amastigotes stage of the parasite replicate in macrophages of the mononuclear phagocyte system and then spread to the entire reticuloendothelial system resulting Kala-azar [27]. Laboratory diagnosis is important for VL because clinical evaluation is not enough for conclusive diagnosis[4]. The sensitivity culture and microscopic examination tends to be low and highly variable, depending on the number of the parasites in sample and technical skills of the personnel [5]. Accurate diagnosis to guide treatment is the first step to achieve the goal of VL elimination. Up until the 1990’s accurate visceral leishmaniasis (VL) diagnosis was complex and invasive including parasitological confirmation by direct microscopic examination of splenic, lymph gland, or bone marrow aspirate or culture of the blood, bone-marrow, lymph nodesor spleen [6]. The rK39 dipstick test is the product of a gene cloned from Leishmania containing 39 amino acid repeat conserved among Leishmania species (7) .A recombinant antigen, rK39 has been shown to be specific for antibodies arising during VL caused by member of the L. donovani complex. It is highly sensitive and predictive for onset of acute disease and evokes high antibody titers of VL patients [8]. The indirect fluorescent antibody (IFA) test is one of the commonly tests used for anti-leishmanial antibody detection by using fixe Promastigotes. The test is based on detecting Ig Gantibodies against Leishmaniaspp, which are demonstrated in the serum very early and different stages of infection but are undetectable up to nine months after cure [9]. Various PCR amplification targets nuclear DNA, such as the small subunit ribosomal RNA gene, internal transcribed spacer regions, the gp63 gene locus and extra chromosomal DNA including the repetitive kinetoplast DNA (kDNA) minicircles [10]. Diagnosis of VL using conventional PCR methods has advanced technique that increasing in sensitivity of assay with optimization of protocols and permitting detection of parasite even before the appearance any clinical symptoms, also applicability to wide variety of clinical material by peripheral blood, lymph node, bone marrow, serum, skin, and urine. Also analysis of archival materials like Giemsa stained bone marrow aspirates, formalin-fixed tissue and skin