Clinical Use of Polymerase Chain Reaction Performed on Peripheral Blood and Bone Marrow Samples for the Diagnosis and Monitoring of Visceral Leishmaniasis in HIV-Infected and HIV-Uninfected Patients: A Single-Center, 8-Year Experience in Italy and Review of the Literature

Antinori, Spinello; Calattini, Sara; Longhi, Erika; Bestetti, Giovanna; Piolini, R.; Magni, C.; Orlando, Giovanna; Gramiccia, Marina; Acquaviva, Verónica; Foschi, Antonella; Corvasce, Stefano; Colomba, Claudia; Titone, Lucina; Parravicini, Carlo; Cascio, Antonio; Corbellino, Mario

doi:10.1086/518167

Cited by 172 publications

(134 citation statements)

References 43 publications

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“…Also, a number of potentially confounding variables were evaluated and controlled for, but multivariable analyses may not entirely eliminate residual confounding from additional unmeasured factors; for example, VL patients had had at least one relapse before inclusion, and findings may be different in patients with a first episode of VL, or without previous relapses. However, previous studies in this population have shown similar results to ours, in terms of immune activation and microbial translocation, independently of the episode [9,29]. Moreover, our results were not related to time of VL or number of relapses.…”

Section: Discussionsupporting

confidence: 87%

“…a concordant response (CR)], and no Leishmania coinfection. Also, 12 healthy control individuals were included in the study for comparison of immunological parameters.A previous diagnosis of VL was defined as a prior parasitological diagnosis by direct visualization of amastigotes in bone marrow and/or peripheral blood, positive Leishmania-specific polymerase chain reaction (PCR) in peripheral blood [29], and clinical symptoms. All VL patients received anti-Leishmania treatment (liposomal amphotericin B, miltefosine or meglumine antimoniate, according to local guidelines) [30], and in all cases they received secondary prophylaxis based on monthly liposomal amphotericin B (3-5 mg/kg) or oral miltefosine (100 mg/daily).…”

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confidence: 99%

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Visceral leishmaniasis as an independent cause of high immune activation, T‐cell senescence, and lack of immune recovery in virologically suppressed HIV‐1‐coinfected patients

Casado¹,

Abad-Fernández²,

Moreno³

et al. 2015

HIV Medicine

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ObjectivesDifferent immune alterations have been described in HIV-infected patients with visceral leishmaniasis (VL). We aimed to identify the immunological factors involved in the lack of immunological recovery and VL relapses in HIV-infected patients with VL, by comparison with other HIV-infected patients. MethodsWe carried out a cross-sectional study of 55 patients receiving suppressive combination antiretroviral therapy (cART) for at least 1 year: nine with previous relapsing VL, 20 with an immunodiscordant response (IDR) to cART (CD4 count < 200 cells/μL) and no previous VL, and 26 with a concordant response (CR) to cART (CD4 count > 350 cells/μL) without VL. Immunosenescence was investigated by analysing CD57 + CD28− levels, immune activation by analysing CD38 + HLA-DR + levels, inflammation by analysing interleukin (IL)-6 levels, and microbial translocation by analysing lipopolysaccharide (LPS) and soluble CD14 (sCD14) levels. ResultsIn VL patients, the median time since VL diagnosis was 42 months, and all patients had had at least one relapse despite suppressive cART for a median time of 43 months. Patients with previously diagnosed VL had a higher CD8 T-cell activation level (P < 0.001) than those with IDR. Also, levels of IL-6, LPS and especially sCD14, associated with bacterial translocation and additional monocyte activation, were significantly increased in patients with previous VL compared with patients with IDR (P = 0.048, P = 0.049 and P < 0.001, respectively). In addition, patients with previous VL had higher levels of CD8 T-cell senescence. Notably, the levels of immune activation and inflammation in patients with previous VL were not related to the time of VL diagnosis, the number of VL relapses, or hepatitis C virus (HCV) coinfection. ConclusionsOur data demonstrate that VL patients had an even worse immunological status than patients with IDR, which was probably associated with increased microbial translocation and additional monocyte/macrophage activation. These data explain the observed lack of immunological recovery and the occurrence of VL relapses in HIV-infected patients with previous VL. Specifically, visceral leishmaniasis (VL) is an infectious disease caused mainly by Leishmania donovani and Leishmania infantum (Leishmania chagasi), intracellular protozoans that infect macrophage cell lineages from lymphoid organs. It is a common coinfection in HIV-1-infected individuals in the Mediterranean basin, including Spain, with a prevalence of up to 9%. VL/HIV-1 coinfection is characterized by significantly lower cure rates, higher drug toxicity, higher relapse rates, and higher mortality rates (nearly 50%) than those for non-HIV-1-infected individuals with VL [5,6].While cART has been shown to decrease the incidence of leishmaniasis in HIV-1-infected patients, it does not seem to prevent the appearance of frequent relapses in patients previously diagnosed and largely pretreated with antiretrovirals [7][8][9], demonstrating the persistence of the parasite in different cell lines. Moreover, ...

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Section: Discussionsupporting

confidence: 87%

mentioning

confidence: 99%

Visceral leishmaniasis as an independent cause of high immune activation, T‐cell senescence, and lack of immune recovery in virologically suppressed HIV‐1‐coinfected patients

Casado¹,

Abad-Fernández²,

Moreno³

et al. 2015

HIV Medicine

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“…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] .…”

Section: Ethical Considerationsmentioning

confidence: 99%

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

Moura

Lopes

Mourão

et al. 2013

Rev. Soc. Bras. Med. Trop.

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Introduction: Rapid diagnostic tests (RDTs) may improve the early detection of visceral leishmaniasis (VL), but their real-world performance requires additional study. Therefore, we evaluated the performance of an rK39-based RDT (Kalazar Detect TM ) for the detection of VL in an endemic, large urban area. Methods: Data were collected from a registry of rK39 RDT performed at 11 emergency care units in Belo Horizonte, Brazil, and from a national database of reportable communicable diseases of the Sistema de Informação de Agravos de Notifi cação (SINAN). Results: The rapid rK39 test was performed in 476 patients, with 114 (23.9%) positive results. The analysis of rK39 RDT performance was based on 381 (80%) cases reported to the SINAN database, of which 145 (38.1%) were confi rmed cases. Estimates for sensitivity and specifi city were 72.4% (95% CI: 64.6-79%) and 99.6% (95%CI: 97.6-99.9%), respectively. Positive and negative predictive values were estimated at 99.1% (95%CI: 94.9-99.8%) and 85.5% (95%CI: 80.8-89.1%), respectively. In addition, close agreement between the rK39 RDT and indirect immunofl uorescence was observed. Conclusions: In summary, the rK39 RDT showed a high specifi city but only moderate sensitivity. In endemic areas for VL, treatment may be considered in cases with clinical manifestations and a positive rK39 RDT, but those with a negative test should be subjected to further investigation.

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Section: Polymerase Chain Reaction (Pcr)mentioning

confidence: 99%

“…The PCR technique has also been used to investigate non-intestinal parasites. The effectiveness of PCR in detecting parasite species such as Leishmania and Plasmodium has already been studied (13)(14)(15)(16)(17)(18)(19)(20)(21)(22).To detect Leishmania in clinical samples, several PCR assays have been developed (17). In studies of visceral Leishmania, PCR has been used for several purposes in addition to diagnosis, such as treatment monitoring and epidemiological studies.…”

mentioning

confidence: 99%

Molecular techniques for the study and diagnosis of parasite infection

Tavares¹,

Staggemeier²,

Borges³

et al. 2011

J. Venom. Anim. Toxins incl. Trop. Dis

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Abstract:In parasitology, routine laboratory diagnosis involves conventional methods, such as optical microscopy, used for the morphological identification of parasites. Currently, molecular biology techniques are increasingly used to diagnose parasite structures in order to enhance the identification and characterization of parasites. The objective of the present study was to review the main current and new diagnostic techniques for confirmation of parasite infections, namely: polymerase chain reaction (PCR), real-time polymerase chain reaction (RT-PCR), loop-mediated isothermal amplification (LAMP), Luminex xMAP, random amplified polymorphic DNA (RAPD), amplified fragment length polymorphism (AFLP), and restriction fragment length polymorphism (RFLP), in addition to microsatellites. Molecular assays have comprehensively assisted in the diagnosis, treatment and epidemiological studies of parasitic diseases that affect people worldwide, helping to control parasitic disease mortality.Key words: parasite infection, diagnosis, molecular techniques, molecular epidemiology. Review ARticle The Journal of Venomous Animals and Toxins including Tropical Diseases ISSN 1678-9199 | 2011 | volume 17 | issue 3 | pages 239-248 INTRODUCTIONIn parasitology, routine laboratory diagnosis involves conventional methods, such as optical microscopy, used for the morphological identification of parasites (1). However, the occasional difficulty of identifying these parasite structures may decrease the sensitivity of such methods. Currently, because of these difficulties, molecular biology has been employed to detect parasites responsible for parasitic diseases (2).Traditional parasitological analyses have the advantage of being less costly without requiring expensive reagents and equipment. Additionally, such analyses can be easily performed when a trained microscopist is available. On the other hand, molecular technology demonstrates the presence of parasites based on their antigenic components or DNA segments (3). These tests are not influenced by environmental factors that usually can interfere with the results of a stool test, for example, thus ensuring highly reliable results (4).Current laboratory diagnostic methods for the identification of parasites include: polymerase chain reaction (PCR), random amplified polymorphic DNA (RAPD), amplified fragment length polymorphism (AFLP), restriction fragment length polymorphism (RFLP), microsatellite marker method, Luminex xMAP-based technology (areas of multianalyte profiling), loop-mediated isothermal amplification (LAMP), and the recently added real-time PCR (5-12).The objective of the present article was to review the applications of molecular biology techniques in parasitology by evaluating and discussing their uses and benefits. MOLECULAR METHODS FOR DETECTION OF PARASITE STRUCTURESSeveral molecular tests to detect parasites have been developed in the last decade. Their specificity and sensitivity have gradually increased, and parasites that were previously difficult to diagnose usi...

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Clinical Use of Polymerase Chain Reaction Performed on Peripheral Blood and Bone Marrow Samples for the Diagnosis and Monitoring of Visceral Leishmaniasis in HIV-Infected and HIV-Uninfected Patients: A Single-Center, 8-Year Experience in Italy and Review of the Literature

Abstract: PCR assay is a highly sensitive and specific tool for the diagnosis of visceral leishmaniasis in both immunocompetent and immunocompromised patients and can be reliably used for rapid parasite identification at the species level.

Cited by 172 publications

References 43 publications

Visceral leishmaniasis as an independent cause of high immune activation, T‐cell senescence, and lack of immune recovery in virologically suppressed HIV‐1‐coinfected patients

Visceral leishmaniasis as an independent cause of high immune activation, T‐cell senescence, and lack of immune recovery in virologically suppressed HIV‐1‐coinfected patients

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

Molecular techniques for the study and diagnosis of parasite infection

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