Comparison of Clinical Samples and Methods in Chronic Cutaneous Leishmaniasis

Eroglu, Fadime; Uzun, Soner; Koltas, Ismail Soner

doi:10.4269/ajtmh.13-0582

Cited by 29 publications

(35 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Articles were excluded at this stage due to (i) articles not available, (ii) not in the field of interest, (iii) use of an inappropriate reference standard, and (iv) inability to complete a 2-by-2 contingency table. A total of 13 articles were included in the systematic review [5,[12][13][14][15][16][17][18][19][20][21][22][23] while data of 12 articles were included in the meta-analysis. Data of 1 article were not considered in the meta-analysis because they did not fit the respective subgroups for molecular method and/or sample type.…”

Section: Included Studiesmentioning

confidence: 99%

Test accuracy of polymerase chain reaction methods against conventional diagnostic techniques for Cutaneous Leishmaniasis (CL) in patients with clinical or epidemiological suspicion of CL: Systematic review and meta-analysis

et al. 2020

View full text Add to dashboard Cite

Background Molecular diagnostic tests, notably polymerase chain reaction (PCR), are highly sensitive test for Leishmania detection, which is especially relevant in chronic cutaneous lesion with lower parasite load. An accurate diagnosis is essential because of the high toxicity of the medications for the disease. Nevertheless, diagnosis of cutaneous leishmaniasis (CL) is hampered by the absence of a reference standard. Assuming that the PCR-based molecular tools are the most accurate diagnostic method, the objective of this systematic review was to assess the diagnostic accuracy of PCR-based molecular tools in a meta-analysis of the published literature. Methodology/Principal findings A search of the published literature found 142 papers of which only 13 studies met the selection criteria, including conventional PCR, real-time PCR, Loop-mediated isothermal amplification (LAMP), recombinase polymerase amplification (RPA), polymorphism-specific PCR (PS-PCR). The sensitivities of the individual studies ranged from 61% to 100%, and specificities ranged from 11% to 100%. The pooled sensitivities of PCR in smears were 0.95 (95% CI, 0.90 to 0.98), and the specificity was 0.91(95% CI, 0.70 to 0.98). In general population, estimates were lower in aspirates, skin biopsies and swab samples with 0.90 (95% CI, 0.80 to 0.95) and 0.87 (95% CI, 0.76 to 0.94) for sensitivity and specificity, respectively. The specificity was lower in consecutive studies, at 0.88 (95% CI, 0.59 to 0.98) and its CI were wider.

show abstract

Section: Included Studiesmentioning

confidence: 99%

Test accuracy of polymerase chain reaction methods against conventional diagnostic techniques for Cutaneous Leishmaniasis (CL) in patients with clinical or epidemiological suspicion of CL: Systematic review and meta-analysis

et al. 2020

View full text Add to dashboard Cite

show abstract

“…It achieves sensitivities and specificities of up to 100% [26,27]. The novel qPCR achieved a lower LOD than cPCR, an outcome seen in other Leishmania assays utilising various targets [14,28,29]. Our future studies will determine the clinical sensitivity of samples previously tested positive for visceral leishmaniasis, to complement the clinical data obtained here.…”

Section: Discussionmentioning

confidence: 65%

Semi-Quantitative, Duplexed qPCR Assay for the Detection of Leishmania spp. Using Bisulphite Conversion Technology

et al. 2019

View full text Add to dashboard Cite

Leishmaniasis is caused by the flagellated protozoan Leishmania, and is a neglected tropical disease (NTD), as defined by the World Health Organisation (WHO). Bisulphite conversion technology converts all genomic material to a simplified form during the lysis step of the nucleic acid extraction process, and increases the efficiency of multiplex quantitative polymerase chain reaction (qPCR) reactions. Through utilization of qPCR real-time probes, in conjunction with bisulphite conversion, a new duplex assay targeting the 18S rDNA gene region was designed to detect all Leishmania species. The assay was validated against previously extracted DNA, from seven quantitated DNA and cell standards for pan-Leishmania analytical sensitivity data, and 67 cutaneous clinical samples for cutaneous clinical sensitivity data. Specificity was evaluated by testing 76 negative clinical samples and 43 bacterial, viral, protozoan and fungal species. The assay was also trialed in a side-by-side experiment against a conventional PCR (cPCR), based on the Internal transcribed spacer region 1 (ITS1 region). Ninety-seven percent of specimens from patients that previously tested positive for Leishmania were positive for Leishmania spp. with the bisulphite conversion assay, and a limit of detection (LOD) of 10 copies per PCR was achieved, while the LOD of the ITS1 methodology was 10 cells/1000 genomic copies per PCR. This method of rapid, accurate and simple detection of Leishmania can lead to improved diagnosis, treatment and public health outcomes.

show abstract

“…Several molecular tests have been used to improve the diagnosis of CL 40,41 ; however, PCR has been found to offer high sensitivity and specificity compared with traditional parasitological methods 37 and can detect parasite DNA in a variety of clinical samples, 6,38 including direct smears, [42][43][44] Flinders Technology Associates filter paper, 45 and swabs from the lesion. 46 In many cases, the efficacy of PCR depends on the DNA extraction process, the number of copies of the molecular marker selected, and the type of PCR used (PCRrestriction fragment length polymorphism, conventional PCR, or qPCR).…”

Section: Discussionmentioning

confidence: 99%

“…In addition, a report by Tiwananthagorn et al 58 concluded that a PCR-kDNA assay was more sensitive than PCR-ITS and LAMP with the 18S rRNA gene. 42 Therefore, a comprehensive study to evaluate these parameters in a vast set of Leishmania species causing CL in the New World is needed.…”

Section: Discussionmentioning

confidence: 99%

Analytical Performance of a Loop-Mediated Isothermal Amplification Assay for Leishmania DNA Detection in Sandflies and Direct Smears of Patients with Cutaneous Leishmaniasis

León

Muñoz

Tabares

et al. 2018

The American Journal of Tropical Medicine and Hygiene

View full text Add to dashboard Cite

Loop-mediated isothermal amplification (LAMP) is ideal for the detection of DNA as it is a quick and easy-to-perform test that does not require complex or sophisticated equipment or infrastructure. However, the application of this technique in the detection of DNA has not been comprehensively analyzed to date (analytical validation). Our objective was to evaluate the sensitivity and analytical specificity (anticipated reportable range [ARR], the limit of detection [LoD], and accuracy) of LAMP targeting the 18S rRNA gene in the diagnosis of six New World species. We then applied the validated LAMP assay across 50 samples of sandflies and 50 direct smears from a recent outbreak of cutaneous leishmaniasis in Colombia to determine its diagnostic performance. The LAMP assay exclusively amplified the DNA of spp., and an ARR of between 1 × 10 and 1 × 10 equivalent parasites/mL was determined. An LoD of 1 × 10 equivalent parasites/mL was established and there was no statistically significant variation in terms of accuracy. Finally, a sensitivity of 100% in direct smears and sandflies samples was calculated and a specificity of 90.9% for direct smears using microscopy as reference and 96.8% for sandflies using real-time polymerase chain reaction as reference were determined. To our knowledge, this is the first attempt to analytically validate a LAMP test to detect DNA, which showed good diagnostic potential from sandflies and direct smear samples.

show abstract

Comparison of Clinical Samples and Methods in Chronic Cutaneous Leishmaniasis

Cited by 29 publications

References 29 publications

Test accuracy of polymerase chain reaction methods against conventional diagnostic techniques for Cutaneous Leishmaniasis (CL) in patients with clinical or epidemiological suspicion of CL: Systematic review and meta-analysis

Test accuracy of polymerase chain reaction methods against conventional diagnostic techniques for Cutaneous Leishmaniasis (CL) in patients with clinical or epidemiological suspicion of CL: Systematic review and meta-analysis

Semi-Quantitative, Duplexed qPCR Assay for the Detection of Leishmania spp. Using Bisulphite Conversion Technology

Analytical Performance of a Loop-Mediated Isothermal Amplification Assay for Leishmania DNA Detection in Sandflies and Direct Smears of Patients with Cutaneous Leishmaniasis

Contact Info

Product

Resources

About