Polymerase Chain Reaction in the Diagnosis and Prognosis of Mediterranean Visceral Leishmaniasis in Immunocompetent Children

Abstract

ABSTRACT. Objective. To assess the usefulness of a polymerase chain reaction (PCR) assay amplifying the small subunit rRNA coding region of Leishmania species performed on peripheral blood (PB) and bone marrow (BM) aspirates for the diagnosis and follow-up of visceral leishmaniasis (VL) in children living in the Mediterranean basin.Design. A prospective study was conducted on children consecutively hospitalized over a 1-year period at our Infectious Diseases Department in Sicily (Italy) presenting with fever, hepatosplenomegaly, and/or pancytopenia and a positive Leishmania serology (>1:40).Results. Among the 14 patients hospitalized with signs and symptoms suggestive of the disease and a positive serology, we identified 10 cases of Mediterranean VL. PCR performed on PB and BM aspirates was positive in all cases and concordant with microscopy and/or culture performed on BM. Leishmania DNA was cleared from PB a median of 6 days after the start of treatment; during follow-up (median: 9 months; range: 6 -12 months) 1 child relapsed. In this case, BM PCR remained positive with rapid reappearance of a positive signal also in PB.Conclusions. PB PCR allows a rapid and noninvasive parasitologic diagnosis of Mediterranean VL among immunocompetent children and is at least as sensitive as a diagnosis made on the basis of BM aspirates. The lack of disappearance from BM and the reappearance of positive PCR on PB is predictive of clinical relapse. Qualitative and semiquantitative PCR may be the standard method for monitoring response to therapy in immunocompetent children. Pediatrics 2002;109(2). URL: http://www. pediatrics.org/cgi/content/full/109/2/e27; visceral leishmaniasis, polymerase chain reaction, diagnosis, peripheral blood, bone marrow.ABBREVIATIONS. VL, visceral leishmaniasis; BMA, bone marrow aspirate; PB, peripheral blood; PCR, polymerase chain reaction; HIV, human immunodeficiency virus; RFLP, restriction fragment length polymorphism; BM, bone marrow. V isceral leishmaniasis (VL) attributable toLeishmania infantum is a vector-borne zoonotic disease transmitted by sand fly bites and is endemic in rural or periurban areas of the Mediterranean basin. 1 Before the age-related changes introduced by the acquired immunodeficiency syndrome epidemic in southern European countries, VL mainly affected children younger than 5 years. 2,3 However, an epidemiologic survey conducted in Sicily between 1987 and 1995 revealed that 53% of all cases were still observed in children younger than 14 years of age. 4 In 2 recent retrospective pediatric studies conducted in France and Greece, the median age of patients was 2 years, 5 months. 5,6 The most common methods of diagnosing VL are classic parasitologic methods such as microscopic direct examination and in vitro culture of bone marrow aspirates (BMAs). As these techniques require invasive procedures and are difficult to repeat for follow-up of patients, an easy, rapid, and noninvasive method would be valuable. In this regard, polymerase chain reaction (PCR) used on peripheral bloo...

Performance of a rapid diagnostic test for the detection of visceral leishmaniasis in a large urban setting

“…Moreover, the high PPV estimates support the World Health Organization 23 and the Brazilian Ministry of Health 7 recommendations for the use of RDTs to confi rm a VL diagnosis in suspected cases. However, due to the moderate estimates for NPV, our fi ndings suggest that patients with negative RDT results should undergo further investigation with additional serological tests and examination of bone marrow aspirates.…”

“…However, due to the moderate estimates for NPV, our fi ndings suggest that patients with negative RDT results should undergo further investigation with additional serological tests and examination of bone marrow aspirates. Polymerase chain reaction for the detection of Leishmania deoxyribonucleic acid (DNA) in bone marrow or peripheral blood specimens might also be useful to confi rm or rule out VL diagnosis in challenging situations [23][24][25][26] .…”

“…In Belo Horizonte, the rK39 RDT was implemented at emergency care units in 2010 to foster the early treatment of suspected VL cases with a positive test, according to World Health Organization (WHO) recommendations 23 . The objective of the present study was to assess rK39 RDT performance in widespread use in a large urban area with a high incidence of VL.…”

Emergence of a new focus of visceral leishmaniasis due to Leishmania infantum in Golestan Province, north-eastern of Iran

“…Based on our previous study that carried out in Fars Province, the sensitivity and specificity of PCR for detection of HVL were determined 82.1 and 100 %, respectively (Fakhar et al 2012a). The PCR technique has several advantages including the ability to work with small amounts of target material, fast detection of Leishmania in symptomatic patients and asymptomatic carriers as well as Leishmania/HIV co-infected patients, the follow up of treatment and the assessment of the successful cure of visceral leishmaniasis (Cascio et al 2002;Bossolasco et al 2003;Riera et al 2004;Maurya et al 2005). It is also useful to complement the serological results (Fakhar et al 2012a).…”

Pediatric visceral leishmaniasis diagnosis in Tunisia: comparative study between optimised PCR assays and parasitological methods

“…Over the last decade, diagnosis has been established by molecular methods. PCR can be performed on any biological sample, bone marrow and blood, and has been applied to routine hospital diagnosis in many laboratories all over the world (Cascio et al, 2002;Lachaud et al, 2000Lachaud et al, , 2001Spanakos et al, 2002). To assess the usefulness of PCR in amplifying Leishmania Genome, we report the results of two optimised PCR assays (R221/R332 and Lei70L/Lei70R) comparing them with classical methods, performed on biological samples, peripheral blood as a less invasive sample compared to bone marrow aspirates.…”