Real-Time PCR Assay for Clinical Management of Human Immunodeficiency Virus-Infected Patients with Visceral Leishmaniasis

Abstract

To evaluate the usefulness of a real-time PCR for Leishmania DNA in the diagnosis and follow-up of patients with human immunodeficiency virus type 1 (HIV-1) and Leishmania coinfection, Leishmania DNA levels were measured in whole peripheral blood from 25 HIV-infected patients with clinical features suggestive of visceral leishmaniasis. Leishmania DNA was detected in 10 of 25 patients with microscopically confirmed visceral leishmaniasis and in none of those without this disease. Following treatment with liposomal amphotericin B, a clinical response was observed in 9 of 10 patients, in association with significantly decreased parasite loads. Seven patients relapsed clinically a median of 110 days after the end of treatment, in association with substantial increases in Leishmania DNA levels. Leishmania DNA levels correlated with the clinical course of visceral leishmaniasis, and their measurement at diagnosis and during and after treatment seems to be useful in the clinical management of HIV-infected patients with this disease.

Diagnosis of visceral leishmaniasis: the sensitivities and specificities of traditional methods and a nested PCR assay

“…Tests based on PCR appear particularly sensitive and allow diagnosis to be based on routine samples of peripheral blood rather than bone-marrow or splenic aspirates (Cascio et al, 2002;Sundar and Rai, 2002;Bossolasco et al, 2003;present study). In adequately equipped laboratories, samples of blood from suspected or treated cases of VL could be routinely screened by PCR, as a relatively non-invasive alternative to the production of smears or cultures of aspirates (Pizzuto et al, 2001;Cascio et al, 2002;Bossolasco et al, 2003). Studies with larger cohorts of patients and in other endemic regions are needed to assess fully the potential usefulness of PCR-based tests.…”

Mediterranean visceral leishmaniasis associated with acute lymphoblastic leukemia (ALL)

“…The PCR technique has several advantages including the ability to work with small amounts of target material, the follow-up of treatment as well as the assessment of the successful cure of visceral leishmaniasis and fast detection of Leishmania in symptomatic patients and asymptomatic carriers as well as Leishmania /HIV co-infected patients (Bossolasco et al 2003;Mary et al 2004;Selvapandiyan et al 2005). The most suitable target for the DNA-based diagnosis is kinetoplast DNA minicircles.…”

Leishmaniasis: diagnostic issues in Europe

“…The drawback of this method is the need for a well-equipped laboratory, while cost still limits its use in developing countries [79]. Two targets for RT-PCR have been investigated: 18S rDNA/small subunit (18S rDNA/SSU) and kinetoplast DNA [77,[80][81][82]. The sensitivity for the 18S rDNA/SSU PCR with whole blood varied from 90 to 100%, while that for the kinetoplast target ranged from 95% with buffy coat cells and 100% with oral fluids [80][81][82].…”

“…Two targets for RT-PCR have been investigated: 18S rDNA/small subunit (18S rDNA/SSU) and kinetoplast DNA [77,[80][81][82]. The sensitivity for the 18S rDNA/SSU PCR with whole blood varied from 90 to 100%, while that for the kinetoplast target ranged from 95% with buffy coat cells and 100% with oral fluids [80][81][82]. Specificities for the 18S rDNA/SSU with whole blood was between 98 and 100%, while that for kinetoplasttargeted PCR was found to be 90% with both buffy coat cells and oral fluids [81,82].…”