scholarly journals Detection of a broad range of human adenoviruses in respiratory tract samples using a sensitive multiplex real-time PCR assay

2008 ◽  
Vol 80 (5) ◽  
pp. 856-865 ◽  
Author(s):  
Sallene Wong ◽  
Kanti Pabbaraju ◽  
Xiaoli L. Pang ◽  
Bonita E. Lee ◽  
Julie D. Fox
2014 ◽  
Vol 20 (12) ◽  
pp. O1010-O1016 ◽  
Author(s):  
S. Bonot ◽  
L. Ogorzaly ◽  
B. El Moualij ◽  
W. Zorzi ◽  
H.-M. Cauchie

2019 ◽  
Vol 64 (8) ◽  
pp. 493-496
Author(s):  
A. S. Pimenova ◽  
O. Yu. Borisova ◽  
M. S. Petrova ◽  
N. T. Gadua ◽  
A. B. Borisova ◽  
...  

The aim of the work was to comparison of rayon and flocked swabs for collection and transport of deep throat swabs for detection of bacteria causing whooping cough by multiplex real-time PCR assay. The study included 87 patients aged from 1 month to 37 years, hospitalized in Infectious Diseases Clinical Hospital No. 1 of the Moscow Department of Healthcare. 68 (78,2 %) people had a diagnosis of whooping cough, the main group of which consisted of children aged 1 to 12 months (median 4 months); 17 (19,5 %) - other diseases of the respiratory tract; 2 (2,3 %) - contact with sick whooping cough. The initial examination of patients was carried out on the 1 - 8th week of the onset of the disease. The material from the patients was taken at one-day interval with commercial rayon swabs and flocked swabs. Identification and differentiation of specific genome fragments of the causative agents of whooping cough in biological material was carried out by real-time PCR using the «AmpliSens® Bordetella multi-FL» reagent kit. The efficiency of PCR-based diagnostics of whooping cough using flocked swabs at the preanalytical stage was 83,8 %, and rayon swabs - 82,3 %. The use of a flocked swabs at the preanalytical stage increased the research efficiency by 1,5 %. Thus, when collecting biological material for PCR-based diagnostics of whooping cough it is possible to use flocked swabs.


2005 ◽  
Vol 71 (6) ◽  
pp. 3131-3136 ◽  
Author(s):  
Narayanan Jothikumar ◽  
Theresa L. Cromeans ◽  
Vincent R. Hill ◽  
Xiaoyan Lu ◽  
Mark D. Sobsey ◽  
...  

ABSTRACT A quantitative real-time TaqMan PCR assay for detection of human adenoviruses (HAdV) was developed using broadly reactive consensus primers and a TaqMan probe targeting a conserved region of the hexon gene. The TaqMan assay correctly identified 56 representative adenovirus prototype strains and field isolates from all six adenovirus species (A to F). Based on infectious units, the TaqMan assay was able to detect as few as 0.4 and 0.004 infectious units of adenovirus serotype 2 (AdV2) and AdV41, respectively, with results obtained in less than 90 min. Using genomic equivalents, the broadly reactive TaqMan assay was able to detect 5 copies of AdV40 (which had zero mismatches with the PCR primers and probe), 8 copies of AdV41, and 350 copies of AdV3 (which had the most mismatches [seven] of any adenovirus serotype tested). For specific detection and identification of F species serotypes AdV40 and AdV41, a second real-time PCR assay was developed using fluorescence resonance energy transfer (FRET) probes that target the adenovirus fiber gene. The FRET-based assay had a detection limit of 3 to 5 copies of AdV40 and AdV41 standard DNA and was able to distinguish between AdV40 and AdV41 based on melting curve analysis. Both the TaqMan and FRET PCR assays were quantitative over a wide range of virus titers. Application of these assays for detection of adenoviruses and type-specific identification of AdV40 and AdV41 will be useful for identifying these viruses in environmental and clinical samples.


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