TY - JOUR
T1 - Development and clinical evaluation of an internally controlled, single-tube multiplex real-time PCR assay for detection of Legionella pneumophila and other Legionella species
AU - Templeton, Kate E.
AU - Scheltinga, Sitha A.
AU - Sillekens, Peter
AU - Crielaard, Jantine W.
AU - van Dam, Alje P.
AU - Goossens, Herman
AU - Claas, Eric C. J.
PY - 2003
Y1 - 2003
N2 - A multiplex real-time PCR assay for detection of Legionella pneumophila and Legionella spp. and including an internal control was designed. Legionella species, L. pneumophila, and the internal control were detected simultaneously by probes labeled with 6-carboxy-fluorescein, hexachlorofluorescein, and indodicarbocyanine, respectively. Therefore, no postamplification analysis was required in order to distinguish the targets. The sensitivity of both assays was 2.5 CFU/ml, and from analysis of 10 culture-positive and 74 culture-negative samples from patients investigated for legionellosis, 100% agreement was observed by both assays in comparison to culture. Four additional positives were found by the multiplex real-time PCR assay in the Legionella culture-negative samples.
AB - A multiplex real-time PCR assay for detection of Legionella pneumophila and Legionella spp. and including an internal control was designed. Legionella species, L. pneumophila, and the internal control were detected simultaneously by probes labeled with 6-carboxy-fluorescein, hexachlorofluorescein, and indodicarbocyanine, respectively. Therefore, no postamplification analysis was required in order to distinguish the targets. The sensitivity of both assays was 2.5 CFU/ml, and from analysis of 10 culture-positive and 74 culture-negative samples from patients investigated for legionellosis, 100% agreement was observed by both assays in comparison to culture. Four additional positives were found by the multiplex real-time PCR assay in the Legionella culture-negative samples.
UR - https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=0041419138&origin=inward
UR - https://www.ncbi.nlm.nih.gov/pubmed/12958219
U2 - https://doi.org/10.1128/JCM.41.9.4016-4021.2003
DO - https://doi.org/10.1128/JCM.41.9.4016-4021.2003
M3 - Article
C2 - 12958219
SN - 0095-1137
VL - 41
SP - 4016
EP - 4021
JO - Journal of clinical microbiology
JF - Journal of clinical microbiology
IS - 9
ER -