Evaluation of urine for Leishmania infantum DNA detection by real-time quantitative PCR

Silva, Rômulo Pessoa e; Trajano-Silva, Lays Adrianne Mendonça; Silva, Maria Almerice Lopes da; Gonçalves-de-Albuquerque, Suênia da Cunha; Goes, Tayná Correia de; Morais, Rayana Carla Silva de; Melo, Fábio Lopes de; Paiva-Cavalcanti, Milena de

doi:10.1016/j.mimet.2016.10.002

Cited by 19 publications

(12 citation statements)

References 40 publications

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“…Of note, diagnostic Leishmania -PCR from urine samples [ 32 , 33 , 34 , 35 ] as well as various associated nucleic acid extraction strategies [ 36 ] have been introduced and recently also summarized in reviews [ 37 , 38 ]. Published interpretations regarding the diagnostic reliability of these approaches vary [ 37 , 38 ].…”

Section: Discussionmentioning

confidence: 99%

Comparison of Three In-House Real PCR Assays Targeting Kinetoplast DNA, the Small Subunit Ribosomal RNA Gene and the Glucose-6-Phosphate Isomerase Gene for the Detection of Leishmania spp. in Human Serum

Tanida

Balczun

Hahn³

et al. 2021

Pathogens

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To perform PCR from serum for the diagnosis of visceral leishmaniasis is convenient and much less invasive than the examination of deeper compartments such as bone marrow. We compared three Leishmania-specific real-time PCRs with three different molecular targets (kinetoplast DNA, the small subunit-ribosomal RNA-(ssrRNA-)gene, the glucose-6-phosphate isomerase-(gpi-)gene) regarding their sensitivity and specificity in human serum. Residual sera from previous diagnostic assessments at the German National Reference Center for Tropical Pathogens Bernhard Nocht Institute for Tropical Medicine Hamburg and the Swiss Tropical and Public Health Institute were used. The sensitivities of kinetoplast DNA-PCR, ssrRNA-gene PCR, and gpi-PCR were 93.3%, 73.3%, and 33.3%, respectively, with 15 initial serum samples from visceral leishmaniasis patients, as well as 9.1%, 9.1%, and 0.0%, respectively, with 11 follow-up serum samples taken at various time points following anti-leishmanial therapy. Specificity was 100.0% in all assays as recorded with 1.137 serum samples from deployed soldiers and migrants without clinical suspicion of visceral leishmaniasis. Kinetoplast-DNA PCR from serum was confirmed as a sensitive and specific approach for the diagnosis of visceral leishmaniasis. The results also indicate the suitability of serum PCR for diagnostic follow-up after therapy, in particular regarding therapeutic failure in case of persisting positive PCR results.

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Section: Discussionmentioning

confidence: 99%

Comparison of Three In-House Real PCR Assays Targeting Kinetoplast DNA, the Small Subunit Ribosomal RNA Gene and the Glucose-6-Phosphate Isomerase Gene for the Detection of Leishmania spp. in Human Serum

Tanida

Balczun

Hahn³

et al. 2021

Pathogens

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“…The diagnostic method works very well for blood samples and even though the number of samples from other sources is reduced, the method borrows our hypothesis of good molecular marker for diagnosis in biological samples with DNA. Amplification was observed even in samples extracted from urine, whose DNA concentration is considerably reduced, corresponding only to fragments of the dead parasite excreted in the urinary system [53]. It should be noted that even with low levels of positivity in the assays using biological samples as a template, we still support our indication of the use of the Cathepsin gene for PCR diagnosis, since even in biological samples where the genetic material of the parasites is scarce or damaged, we obtained satisfactory amplifications.…”

Section: Discussionmentioning

confidence: 99%

Exploring Leishmania infantum cathepsin as a new molecular marker for phylogenetic relationships and visceral leishmaniasis diagnosis

et al. 2019

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BackgroundLeishmania infantum, the etiological agent of visceral leishmaniasis, is a neglected zoonosis that requires validation and standardization of satisfactory diagnostic methodologies. Thus, the aim of the present study was to evaluate the effectiveness of cathepsin L-like protease as a target for making molecular diagnoses and as a phylogenetic marker enabling to understand the intraspecies variations and evolutionary history of L. infantum in Brazil.MethodsWe used 44 isolates of L. infantum. The cathepsin L-like gene fragments were amplified, sequenced, manually aligned and analyzed using inference methods. The sequences generated were used to search and design oligonucleotide primers to be used in reactions specific to the target parasite.ResultsThe cathepsin L-like gene did not show any intraspecies variability among the isolates analyzed. The pair of primers proposed amplified the target deoxyribonucleic acid (DNA) of L. infantum isolates and were effective for DNA amplification at concentrations of as low as 10− 11 ng/μl. The proposed marker did not present cross-reactions with other hemoparasites. When used for making the diagnosis in a panel of clinical samples from dogs, a positivity rate of 49.03% (102/208) was obtained, versus 14.42% (30/208) for a ribosomal internal transcribed spacer (ITS) marker. In samples from sandflies, the rate was 6.25% and from humans, 14.28%.ConclusionsThe results described in this work allow us to infer that CatLeish-PCR is a sensitive and specific marker for use in diagnostic trials of L. infantum and in clinical and epidemiological surveys.

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“…However, egg detection method is below optimum and requires multiple samples; other methods for the detection of parasite in urine are being investigated. PCR shows promising results in this regards, and many papers are advocating for the clinical establishment of this method for detection of parasites such as S. haematobium, Leishmania infantum, Trypanosoma sp., and others [59][60][61][62][63][64].…”

Section: Parasite Detection In Urinementioning

confidence: 98%

Urine Tests for Diagnosis of Infectious Diseases and Antibiotic-Resistant Pathogens

Eltai¹,

Alhussain²,

Doiphode³

et al. 2020

Pathogenic Bacteria

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The relation between disease and urine was recognized by physicians since the earliest civilization BC. Urine is considered an ideal diagnostic specimen for its noninvasive and easy method of collection. Urinalysis encompasses a wide range of tests, which includes a variety of chemical tests, urine microscopy, bacterial cultures, and molecular tests. Importantly, urine tests can diagnose patients with antibiotic-resistant urinary tract infections (UTI), directly from urine and/or bacterial culture. This chapter summarizes the most common urine tests in the infectious disease field, with a special focus on diagnosing UTI and characterizing their antibiotic resistant. In addition to describing the advantages and limitation of these tests, the chapter explores the promising emerging technologies and methods in this field. This chapter is beneficial for scientists and healthcare workers in the field.

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Evaluation of urine for Leishmania infantum DNA detection by real-time quantitative PCR

Cited by 19 publications

References 40 publications

Comparison of Three In-House Real PCR Assays Targeting Kinetoplast DNA, the Small Subunit Ribosomal RNA Gene and the Glucose-6-Phosphate Isomerase Gene for the Detection of Leishmania spp. in Human Serum

Comparison of Three In-House Real PCR Assays Targeting Kinetoplast DNA, the Small Subunit Ribosomal RNA Gene and the Glucose-6-Phosphate Isomerase Gene for the Detection of Leishmania spp. in Human Serum

Exploring Leishmania infantum cathepsin as a new molecular marker for phylogenetic relationships and visceral leishmaniasis diagnosis

Urine Tests for Diagnosis of Infectious Diseases and Antibiotic-Resistant Pathogens

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