Development and Validation of a PCR-ELISA for the Diagnosis of Symptomatic and Asymptomatic Infection by<i>Leishmania (Leishmania) infantum</i>

Medeiros, Fernanda Alvarenga Cardoso; Gomes, Luciana I.; Oliveira, Edward; Souza, Carolina Senra Alves de; Mourão, Maria Vitória Assumpção; Cota, Gláucia; Marques, Letícia Helena dos Santos; Carneiro, Mariângela; Rabello, Ana

doi:10.1155/2017/7364854

Cited by 15 publications

(15 citation statements)

References 46 publications

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“…Similarly, the diagnosis of asymptomatic infection in humans is also challenging because the time between infection development and production of antibodies at detectable levels is not clear, providing room for molecular techniques for enhancing the early detection of infection. 42 , 43 …”

Section: Discussionmentioning

confidence: 99%

Improving the reference standard for the diagnosis of canine visceral leishmaniasis: a challenge for current and future tests

Teixeira

Silva

Vital

et al. 2019

Mem. Inst. Oswaldo Cruz

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BACKGROUND Studies aimed at validating canine visceral leishmaniasis diagnostic tests present heterogeneous results regarding test accuracy, partly due to divergences in reference standards used and different infection evolution periods in animals. OBJECTIVE This study aimed to evaluate the accuracy of the rapid test-dual path platform (TR-DPP) (Biomanguinhos®), EIE-Leishmaniose-Visceral-Canina-Biomanguinhos (EIE-LVC) (Biomanguinhos®), enzyme-linked immunosorbent assay (ELISA) rK39 (in-house), and the direct agglutination test (DAT- Canis ) against a reference standard comprising parasitological and molecular techniques. METHODS A phase II/III validation study was carried out in sample sera from 123 predominantly asymptomatic dogs living in an area endemic for visceral leishmaniasis. FINDINGS Sixty-nine (56.1%) animals were considered infected according to the reference standard. For each test, the sensitivity and specificity, respectively, were as follows: TR-DPP, 21.74% [confidence interval (CI)95% 13.64% to 32.82%] and 92.59% (CI95% 82.45% to 97.08%); EIE-LVC, 11.59% (CI95% 5.9% to 21.25%) and 90.74% (CI95% 80.09% to 95.98%); ELISA rK39, 37.68% (CI95% 27.18% to 49.48%) and 83.33% (CI95% 71.26% to 90.98%); and DAT- Canis , 18.84% (CI95% 11.35% to 29.61%) and 96.30% (CI95% 87.46% to 98.98%). CONCLUSION We concluded that improving the sensitivity of serum testing for diagnosing asymptomatic dogs must constitute a priority in the process of developing new diagnostic tests to be used in the visceral leishmaniasis control program in Brazil.

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Section: Discussionmentioning

confidence: 99%

Improving the reference standard for the diagnosis of canine visceral leishmaniasis: a challenge for current and future tests

Teixeira

Silva

Vital

et al. 2019

Mem. Inst. Oswaldo Cruz

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“…Additionally, the heightened anti-leishmanial IgG and its subclasses have helped identify active VL, and furthermore, reduction of IgG1 titres was associated with successful treatment ( Saha et al., 2005 ; Ansari et al., 2008 ; Bhattacharyya et al., 2014 ). However, the ICT/ELISA technique failed to detect HIV/VL co-infected cases, possibly owing to their immunosuppressed status ( Medeiros et al., 2017 ). Another limitation was that the ICT showed positivity in a significant proportion of apparently healthy individuals in endemic regions as also remained positive for long periods after cure from VL ( Das et al., 2020 ).…”

Section: Resultsmentioning

confidence: 99%

Molecular Tracking of the Leishmania Parasite

Moulik¹,

Sengupta²,

Chatterjee³

2021

Front. Cell. Infect. Microbiol.

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With the Visceral Leishmaniasis/Kala-azar Elimination Program in South Asia in its consolidation phase, the focus is mainly on case detection, vector control, and identifying potential sources of infection. Accordingly, emphasis is presently on curbing transmission, which is potentially achievable by identification and elimination of potential reservoirs. The strongest contenders for being the disease reservoir are cases of Post Kala-azar Dermal Leishmaniasis (PKDL) which occurs in a minor proportion of individuals apparently cured of Visceral Leishmaniasis (VL). The demonstration of parasites in tissue aspirates despite being a risky and invasive process is the gold standard for diagnosis of VL, but is now being replaced by serological tests e.g., rK39 strip test and direct agglutination test. However, these antibody based tests are limited in their ability to diagnose relapses, detect cases of PKDL, and monitor effectiveness of treatment. Accordingly, detection of antigen or nucleic acids by polymerase chain reaction has been successfully applied for monitoring of parasite kinetics. This review article provides updated information on recent developments regarding the available antibody or antigen/nucleic acid based biomarkers for longitudinal monitoring of patients with VL or PKDL and emphasizes the need for availability of studies pertaining to quantification of treatment response or relapse.

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“…Even if the qPCR with the SYBR Green system is considered promising for the diagnosis of asymptomatic infection by L. infantum , no approach can, at the moment, be regarded as a gold standard (Kaushal et al ., 2017; Varani et al ., 2017; Galluzzi et al ., 2018). The ‘best approach’ depends on factors such as assay design, the selected target region, origin of the clinical sample, cycle temperature and the DNA extraction method (Gomes et al ., 2007; Medeiros et al ., 2017).…”

Section: Discussionmentioning

confidence: 99%

Identification of Leishmania infantum in blood donors from endemic regions for visceral leishmaniasis

et al. 2020

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Visceral leishmaniasis is an endemic protozoonosis observed in over 60 countries, with over 500 000 new cases recorded annually. Although the diagnostic procedure of its symptomatic forms is well established, for asymptomatic patients, who represent about 85% of those infected, there is no consensus on the best method for its identification. Recent studies have presented molecular techniques as viable identification methods, with good sensitivity and specificity indices in asymptomatic individuals. Therefore, we aimed to use molecular methods to assess their effectiveness in identifying the presence of asymptomatic infection by Leishmania infantum (L. infantum) individuals from endemic regions of Brazil. Screening was performed by real-time polymerase chain reaction (qPCR) and confirmed by sequencing the cytochrome B gene. Of the 127 samples [from 608 blood donors who had participated in a previous study, of which 34 were positive by the enzyme-linked immunosorbent assay (ELISA) rK39] tested by qPCR, 31 (24.4%) were positive. In the sequencing of 10 qPCR-positive samples, five were identified as L. infantum. Complimentary samples of the ELISA rK39 and conventional PCR showed only reasonable and low agreement with qPCR, respectively. The qPCR confirmed the presence of infection in five of the 10 sequenced samples, ELISA confirmed three, and the conventional PCR confirmed none.

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Development and Validation of a PCR-ELISA for the Diagnosis of Symptomatic and Asymptomatic Infection byLeishmania (Leishmania) infantum

Cited by 15 publications

References 46 publications

Improving the reference standard for the diagnosis of canine visceral leishmaniasis: a challenge for current and future tests

Improving the reference standard for the diagnosis of canine visceral leishmaniasis: a challenge for current and future tests

Molecular Tracking of the Leishmania Parasite

Identification of Leishmania infantum in blood donors from endemic regions for visceral leishmaniasis

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