Leishmania parasite detection and quantification using PCR-ELISA

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216208%3A11310%2F10%3A10082049" target="_blank" >RIV/00216208:11310/10:10082049 - isvavai.cz</a>

Alternative codes found

RIV/68378050:_____/10:00355298

Result on the web

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DOI - Digital Object Identifier

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Result language

angličtina

Original language name

Leishmania parasite detection and quantification using PCR-ELISA

Original language description

This protocol describes an improved and optimized PCR-ELISA method for detection and quantification of Leishmania parasites in host tissues. Unlike other DNA-based assays, this method uses digoxigenin-and biotin-labeled primers. This eliminates the needfor a separate step of hybridization of the PCR product with labeled probes. The PCR product is detected using sandwich ELISA with antidigoxigenin-detecting antibodies. Primers are complementary to the kinetoplast minicircle conserved region of parasiteDNA, allowing the detection of several Leishmania species. For measurement of a wide range of parasite concentrations, +/- 25 cycles were optimal. the sensitivity of this technique is 0.3 fg of parasite DNA per reaction in 40-cycle PCR-ELISA, corresponding to 0.004 parasites. DNA preparation by a standard TRI reagent procedure takes about 4 h. When DNA is prepared, a single person can test a large number of samples (at least 150) in a maximum of 7 h.

Czech name

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Czech description

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Type

J_x - Unclassified - Peer-reviewed scientific article (Jimp, Jsc and Jost)

CEP classification

EB - Genetics and molecular biology

OECD FORD branch

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Project

Result was created during the realization of more than one project. More information in the Projects tab.

Continuities

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Publication year

2010

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

Nature Protocols

ISSN

1754-2189

e-ISSN

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Volume of the periodical

5

Issue of the periodical within the volume

6

Country of publishing house

GB - UNITED KINGDOM

Number of pages

7

Pages from-to

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UT code for WoS article

000278354700010

EID of the result in the Scopus database

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