Linked Life Data

10974143

Source:http://linkedlifedata.com/resource/pubmed/id/10974143

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
4
pubmed:dateCreated
2000-10-19
pubmed:abstractText
We developed a nucleic acid sequence based amplification (NASBA) assay which employs the recombinant photoprotein Aequorin in a microtiter plate format for detection and quantitation of C. trachomatis that may be useful in large scale epidemiological studies aimed at improving our understanding of factors affecting transmission of this sexually transmitted pathogen. The conditions for NASBA amplification of the16S rRNA target were optimized (90 mM KCl, 12 mM MgCl(2), 0.2 microM P1 and P2 primers), amplified RNA was captured by a biotin-labelled capture probe immobilized onto streptavidin coated microtiter plates and detected with an FITC-labelled oligonucleotide probe and Aequorin-anti-FITC antibody conjugate. The analytical sensitivity of NASBA was 1,000 in vitro generated RNA transcripts and 1.6 IFU of C. trachomatis. The sensitivity of NASBA using the bioluminescent assay was 10 fold higher than Northern blotting. Time course amplification experiments performed with 10 fold serial dilutions of target established that amplification was linear at 75 min and extended over a range of five log units of input RNA copy number. Linear regression analysis confirmed a linear fit for the data with r(2) = 0.959 (p < 0.004). A double log plot of RLU signal versus copy number was linear; analysis of residuals from a series of runs tests confirmed a fit with a linear model (number of runs = 3, p = 0.5 where p < 0.05 indicates statistical deviation from a linear model). NASBA amplification coupled with bioluminescent detection in a microtiter plate format should provide a useful tool for quantitation of C. trachomatis in clinical specimens for use in epidemiological studies.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Aug
pubmed:issn
1386-2073
pubmed:author
pubmed:issnType
Print
pubmed:volume
3
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
303-13
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed:year
2000
pubmed:articleTitle
Quantitation of Chlamydia trachomatis 16S rRNA using NASBA amplification and a bioluminescent microtiter plate assay.
pubmed:affiliation
The Father Sean O- Sullivan Research Center, St. Joseph-s Hospital, McMaster University, Hamilton, L8N 4A6, CANADA.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't