Application of PCR-ELISA in Molecular Diagnosis

Sue, Mei Jean; Yeap, Swee Keong; Omar, Abdul Rahman; Tan, Sheau Wei

doi:10.1155/2014/653014

Cited by 47 publications

(28 citation statements)

References 47 publications

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“…This method combines PCR and ELISA into a single analytical technique and provides an objective interpretation of the results compared to conventional PCR followed by electrophoresis because it uses a gene-specific probe to detect the amplified product. The PCR-ELISA also allows multiple sample testing and quantitative and qualitative analyses, such as quantitative real-time PCR (qRT-PCR), but using basic laboratory equipment for the processing ELISA tests that is present in most clinical laboratories [20, 21, 24]. PCR-ELISA has the additional advantage of using a whole blood sample that is obtained in a minimally invasive manner.…”

Section: Discussionmentioning

confidence: 99%

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

Medeiros

Gomes

Oliveira

et al. 2017

Journal of Tropical Medicine

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A kDNA PCR enzyme-linked immunosorbent assay (kDNA PCR-ELISA) for the diagnosis of human visceral leishmaniasis (HVL) was developed. The detection limit of the reaction, precision measurements, and cut-off of the kDNA PCR-ELISA were defined in a proof-of-concept phase. A reference strain of Leishmania (Leishmania) infantum and a bank of 14 peripheral blood samples from immunocompetent patients with VL were characterized using techniques considered gold standards, and 11 blood samples obtained from healthy individuals of an endemic area were also assessed. Phase II evaluation determined the performance of the assay in peripheral blood samples from 105 patients with VL (adults and children), 25 patients with Leishmania/HIV coinfection, 40 healthy individuals, and 33 asymptomatic individuals living in endemic areas. The kDNA PCR-ELISA exhibited satisfactory precision, with a detection limit of 0.07 fg of DNA from L. (L.) infantum and 1 parasite/mL blood. The overall sensitivity of the assay for all groups studied was 100% (95% confidence interval [CI]: 97.1–100%), and the specificity was 95% (95% CI: 83.5–98.6%). The kDNA PCR-ELISA was shown to be a useful tool for VL symptomatic and asymptomatic individuals diagnosis and its use in endemic countries may help monitor control interventions.

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Section: Discussionmentioning

confidence: 99%

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

Medeiros

Gomes

Oliveira

et al. 2017

Journal of Tropical Medicine

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“…The PCR-ELISA method detects nucleic acid instead of protein and is much more sensitive compared to conventional PCR assays, with lower detection limit and shorter analytical time (30). The most basic step in designing a diagnostic reaction is the selection of suitable fragments and designing necessary primers for the purpose of segments duplication.…”

Section: Discussionmentioning

confidence: 99%

Rapid and Specific Polymerase Chain Reaction-Enzyme Linked Immunosorbent Assay for Detection of Escherichia coli LT Toxin From Clinical Isolates

Esfandiari

Amani

Fouladi

et al. 2016

Arch Clin Infect Dis

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Background: Enterotoxigenic Escherichia coli (ETEC) is the most common agent, which causes diarrhea. ETEC is colonized along the cells and produces enterotoxins leading to diarrhea. Different detection methods have been utilized for detection of ETEC heat Labile Toxin (LT) toxins or respective genes. These methods have disadvantages such as high costs and labor time and limitations in handling many samples simultaneously.

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“…A number of researchers are using PCR-ELISA for early detection and quantification which allows multiple sample testing using whole blood with sensitivity of 87% [101, 147]. However this method is tedious, expensive, and less sensitive than qPCR and tested on limited number of clinical samples [101, 148]. …”

Section: Standard Diagnostic Tools and Their Limitationsmentioning

confidence: 99%

Molecular Diagnosis of Visceral Leishmaniasis

Sundar

2018

Mol Diagn Ther

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Visceral leishmaniasis (VL), a deadly parasitic disease, is a major public health concern globally. Countries affected by VL have signed the London Declaration on Neglected Tropical Diseases and committed to eliminate VL as a public health problem by 2020. To achieve and sustain VL elimination, it will become progressively important not to miss any remaining cases in the community who can maintain transmission. This requires accurate identification of symptomatic and asymptomatic carriers using highly sensitive diagnostic tools at the primary health service setting. The rK39 rapid diagnostic test (RDT) is the most widely used tool and with its good sensitivity and specificity is the first choice for decentralized diagnosis of VL in endemic areas. However, this test cannot discriminate between current, subclinical, or past infections and is useless for diagnosis of relapses and as a prognostic (cure) test. Importantly, as the goal of elimination of VL as a public health problem is approaching, the number of people susceptible to infection will increase. Therefore, correct diagnosis using a highly sensitive diagnostic test is crucial for applying appropriate treatment and management of cases. Recent advances in molecular techniques have improved Leishmania detection and quantification, and therefore this technology has become increasingly relevant due to its possible application in a variety of clinical sample types. Most importantly, given current problems in identifying asymptomatic individuals because of poor correlation between the main methods of detection, molecular tests are valuable for VL elimination programs, especially to monitor changes in burden of infection in specific communities. This review provides a comprehensive overview of the available VL diagnostics and discusses the usefulness of molecular methods in the diagnosis, quantification, and species differentiation as well as their clinical applications.

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Application of PCR-ELISA in Molecular Diagnosis

Cited by 47 publications

References 47 publications

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

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

Rapid and Specific Polymerase Chain Reaction-Enzyme Linked Immunosorbent Assay for Detection of Escherichia coli LT Toxin From Clinical Isolates

Molecular Diagnosis of Visceral Leishmaniasis

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