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A group of 76 consecutive human immunodeficiency virus (HIV)-positive patients with fever of unknown origin (n ‫؍‬ 52) or fever associated with pulmonary diseases was evaluated in order to assess the usefulness of PCR with peripheral blood in the diagnosis and follow-up of visceral leishmaniasis. We identified 10 cases of visceral leishmaniasis among the 52 patients with fever of unknown origin. At the time of diagnosis, all were parasitemic by PCR with peripheral blood. During follow-up, a progressive decline in parasitemia was observed under therapy, and all patients became PCR negative after a median of 5 weeks (range, 6 to 21 weeks). However, in eight of nine patients monitored for a median period of 88 weeks (range, 33 to 110 weeks), visceral leishmaniasis relapsed, with positive results by PCR with peripheral blood reappearing 1 to 2 weeks before the clinical onset of disease. Eight Leishmania infantum and two Leishmania donovani infections were identified by PCR-restriction fragment length polymorphism analysis. PCR with peripheral blood is a reliable method for diagnosis of visceral leishmaniasis in HIV-infected patients. During follow-up, it substantially reduces the need for traditional invasive tests to assess parasitological response, while a positive PCR result is predictive of clinical relapse.

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...

Summary Objectives: To analyse the risk determinants of HIV-1 and HIV-2 infections in pregnant women in Bissau. Methods: Pregnant women attending the antenatal clinics of Bissau between January 2002 and June 2006 were consecutively tested unless they opted out. Results: Among 23,869 tested women the overall prevalence of HIV-1 was 5.7%, that of HIV-2 was 2.4%, and these included the 0.7% prevalence of HIV-1 and HIV-2 duals. The main factors associated with the risk of HIV-1 infection were older age, occupation and number of sexual partners. Beafada and Mandinga ethnic groups were at greater risk of presenting HIV-1, and Bijago and Papel at lower risk. The factors associated with the risk of HIV-2 were age, literacy and occupation; the Beafada were at greater risk than the other ethnic groups. Conclusions: The prevalence of HIV-2 infection decreased overtime, whereas that of HIV-1 infection remained substantially stable, but was higher than that observed in previous studies. The rapid decline in the rates of HIV-2 infection suggests that many of the factors that allowed its exponential growth in the past have now been partially removed, and that sexual and vertical transmission have not been sufficient to maintain and extend the epidemic.