A urine-based polymerase chain reaction method for the diagnosis of visceral leishmaniasis in immunocompetent patients

Fakhar, Mehdi; Motazedian, Mohammad Hossein; Hatam, Gholamreza; Mikaeili, Fattaneh

doi:10.1016/j.diagmicrobio.2007.09.001

Cited by 47 publications

(24 citation statements)

References 34 publications

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“…Leishmania may be cultured on a Novy-McNeal Nicolle medium but this is not done routinely in several countries due to the required expertise and cost. Despite lacking standardisation conserved sequences in minicircle kinetoplast DNA or in the small subunit rRNA gene of Leishmania may be detected rapidly by PCR on lymph node and bone marrow aspirates (Case 7), on peripheral blood 31 or on urine 32 …”

Section: Discussionmentioning

confidence: 99%

Manifestations of paediatric Leishmania infantum infections in Malta

Pace

Williams

Grochowska

et al. 2011

Travel Medicine and Infectious Disease

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SummaryLeishmania infantum is endemic in the Maltese archipelago, a group of islands in the Mediterranean which are visited frequently by tourists from Northern European countries. The burden of leishmaniasis is highest in children who may present with cutaneous or visceral manifestations. We describe systematically the manifestations, diagnosis and management of leishmaniasis in children <14 years of age, who had a histopathological diagnosis of leishmaniasis in Malta, from 2004 to 2008. Eleven children were diagnosed with leishmaniasis; 8 children (15–44 months of age) had visceral disease and three (aged 9–13 years) suffered cutaneous infections. Prolonged high grade fever, pallor, hepatosplenomegaly, and pancytopenia were common presenting features of visceralisation. Diagnosis was based on the visualisation of amastigotes from bone marrow aspirates. Pentavalent antimonials were associated with treatment failure in two children, whilst liposomal amphotericin B was curative in all. Children with cutaneous leishmaniasis had dry crusted ulcero-nodular lesions on exposed areas which responded to intra-lesional instillation of sodium stibogluconate or to cryotherapy. Leishmaniasis should be included in the differential diagnosis of fever and hepatosplenomegaly or chronic cutaneous lesions in children who travel to Malta.

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Section: Discussionmentioning

confidence: 99%

Manifestations of paediatric Leishmania infantum infections in Malta

Pace

Williams

Grochowska

et al. 2011

Travel Medicine and Infectious Disease

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“…Attempts have been made to develop urine/conjunctival swab-based PCR for ease of sample collection particularly in infants and to minimize the risk of blood-borne infections (Motazedian et al 2008;Pilatti et al 2009). …”

Section: Polymerase Chain Reactionmentioning

confidence: 99%

Diagnosis of visceral leishmaniasis: developments over the last decade

et al. 2011

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Diagnostic parameters for visceral leishmaniasis (VL), a potentially fatal parasitic disease caused by Leishmania donovani, have been redefined in the last decade with the development of serological and molecular tests, though a definitive diagnosis still banks on the century-old parasitological methods in many areas. Recombinant antigens have improved performance of serodiagnostic methods. Serology-based tests, rk39 antigen dipstick, and direct agglutination test commonly employed in the field are highly sensitive methods, however, fail to distinguish past infections. Molecular approaches have become increasingly relevant due to remarkable sensitivity, specificity, and flexibility in choice of samples. Quantitative polymerase chain reaction is a highly sensitive and specific tool used in referral labs for detection/assessment of parasite load in VL patients and subsequently in monitoring treatment response to antileishmanial agents. The method displays potential to provide threshold for distinguishing asymptomatics in endemic areas. Currently, improvement in VL diagnostics is required for successful decentralized (point-of-care) testing in field conditions and to detect VL-HIV co-infection. Techniques such as loop-mediated isothermal amplification offer a reliable molecular diagnostic method for field application. The diagnosis based on bioanalytics/biosensors promise frontiers for point-of-care VL detection after adequate standardization. This review summarizes the recent developments in VL diagnostics, drawing attention towards the need for standardization of the diagnostics across the affected regions.

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“…PCR assays has been also performed in non invasive samples like buccal swab and urine with sensitivity of 83% and 88% respectively [95, 96]. Molecular diagnosis using PCR is very useful in HIV-VL patients where clinical picture is confusing and, serological as well as immunological tests are not reliable due to low sensitivity [97]. Furthermore, sensitivity and specificity of PCR for detection of low level parasitemia has been shown to be significantly improved by performing nested and semi nested PCR which involves two sets of primers (targeting single gene locus) used in two successive runs.…”

Section: Standard Diagnostic Tools and Their Limitationsmentioning

confidence: 99%

Molecular Diagnosis of Visceral Leishmaniasis

Sundar

2018

Mol Diagn Ther

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Visceral leishmaniasis (VL), a deadly parasitic disease, is a major public health concern globally. Countries affected by VL have signed the London Declaration on Neglected Tropical Diseases and committed to eliminate VL as a public health problem by 2020. To achieve and sustain VL elimination, it will become progressively important not to miss any remaining cases in the community who can maintain transmission. This requires accurate identification of symptomatic and asymptomatic carriers using highly sensitive diagnostic tools at the primary health service setting. The rK39 rapid diagnostic test (RDT) is the most widely used tool and with its good sensitivity and specificity is the first choice for decentralized diagnosis of VL in endemic areas. However, this test cannot discriminate between current, subclinical, or past infections and is useless for diagnosis of relapses and as a prognostic (cure) test. Importantly, as the goal of elimination of VL as a public health problem is approaching, the number of people susceptible to infection will increase. Therefore, correct diagnosis using a highly sensitive diagnostic test is crucial for applying appropriate treatment and management of cases. Recent advances in molecular techniques have improved Leishmania detection and quantification, and therefore this technology has become increasingly relevant due to its possible application in a variety of clinical sample types. Most importantly, given current problems in identifying asymptomatic individuals because of poor correlation between the main methods of detection, molecular tests are valuable for VL elimination programs, especially to monitor changes in burden of infection in specific communities. This review provides a comprehensive overview of the available VL diagnostics and discusses the usefulness of molecular methods in the diagnosis, quantification, and species differentiation as well as their clinical applications.

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A urine-based polymerase chain reaction method for the diagnosis of visceral leishmaniasis in immunocompetent patients

Cited by 47 publications

References 34 publications

Manifestations of paediatric Leishmania infantum infections in Malta

Manifestations of paediatric Leishmania infantum infections in Malta

Diagnosis of visceral leishmaniasis: developments over the last decade

Molecular Diagnosis of Visceral Leishmaniasis

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