Could kDNA-PCR in Peripheral Blood Replace the Examination of Bone Marrow for the Diagnosis of Visceral Leishmaniasis?

Abstract

The aim of this study was to evaluate whether the molecular (kDNA-PCR) and parasitological diagnosis in peripheral blood (PB) could replace the invasive and painful bone marrow collection (BM) in the diagnosis of visceral leishmaniasis (VL). PB from suspected VL patients was evaluated by parasitological and molecular techniques using as the gold standard (GS) a combination of clinical, epidemiological, and immunochromatographic test (PB-rK39) results and parasitological examination of BM. Based on the GS, 38 samples from 32 patients were grouped: Group 1, 20 samples of VL cases, and Group 2, 18 samples of non-VL cases. In order to evaluate the parasitological and molecular techniques in PB, the samples were examined. From Group 1, PB kDNA-PCR was positive in 20 samples and in 19 of 20 in BM kDNA-PCR examination. However, the parasitological examination of buffy coat was insensitive, being able to detect only 4 cases from Group 1. All samples from Group 2 were negative. We concluded that, for the diagnosis of visceral leishmaniasis, the parasitological examination of peripheral blood was not useful; however, molecular diagnosis by kDNA-PCR, performed in peripheral blood, could be useful to replace the parasitological examination of bone marrow.

Serological and molecular tools to diagnose visceral leishmaniasis: 2-years’ experience of a single center in Northern Italy

“…VL diagnosis was performed by a combination of clinical and parasitological data; the association of serological and molecular tools has become, in reference centers, a common approach to the differential diagnosis of VL, because of serious prognostic implications of an incorrect or late diagnosis of VL (as reviewed in [ 20 ]). Over the last decade, nucleic acid testing by PCR has emerged as a highly sensitive and specific diagnostic method to detect leishmanial DNA [ 13 , 14 , 20 – 24 ]. Various PCR assays have been validated for the diagnosis of VL, demonstrating higher sensitivity compared to microscopy [ 13 , 22 – 24 ].…”

“…Over the last decade, nucleic acid testing by PCR has emerged as a highly sensitive and specific diagnostic method to detect leishmanial DNA [ 13 , 14 , 20 – 24 ]. Various PCR assays have been validated for the diagnosis of VL, demonstrating higher sensitivity compared to microscopy [ 13 , 22 – 24 ]. In addition, studies performed in dogs indicate that PCR can identify oligosymptomatic infections caused by Leishmania [ 25 – 27 ]; similarly molecular tools may contribute to detect Leishmania infection in patients presenting with atypical symptoms or with paucisymptomatic infection.…”

Mathematical models of amino acid panel for assisting diagnosis of children acute leukemia

“…The diagnosis of AL is dependent on the multiple laboratory tests, which require the combination of assays of morphological, immunological, cytogenetic and molecular (MICM) inputs [2]. The current procedure (MICM assays) of using bone marrow cells from AL patients is painful and inconvenient for children [3]. The immunological tests rely on flow cytometry, while the molecular tests, such as, reverse transcription polymerase reaction (RT-PCR) and high throughput sequencing are used to measure fusion genes and key mutations of the driven genes.…”

Determination of genetic diversity of Leishmania species using mini-circle kDNA, in Iran-Iraq countries border

“…[15] Kinetoplast is a unique DNA containing structure in the mitochondrion of the cell, consisting of two components, maxi-circle (present in a number of 30–50 copies/parasite, with 20–40 kb in length) and mini-circle kDNA (present in a number of 10,000–20,000 copies/parasite with 1 kb in length). [16171819] Kinetoplast DNA minicircles are often the first choice for leishmaniasis diagnosis, containing three highly conserved blocks conserved sequence blocks (CSB) (CSB-1, conserved blocks of synteny-2 and CSB-3). [20] The main advantage of minicircles is high copy number and variation of kDNA mini-circles, used for the analysis of genetic polymorphism.…”

“…[20] The main advantage of minicircles is high copy number and variation of kDNA mini-circles, used for the analysis of genetic polymorphism. [4161721] Therefore, it is an ideal target for the identification of Leishmania species. [22] Hence, we used kDNA-PCR to determine the genetic diversity of this protozoon, in Ilam Province.…”