scholarly journals Multiplex real-time quantitative PCR, microscopy and rapid diagnostic immuno-chromatographic tests for the detection of Plasmodium spp: performance, limit of detection analysis and quality assurance

2009 ◽  
Vol 8 (1) ◽  
pp. 284 ◽  
Author(s):  
Krishna Khairnar ◽  
Donald Martin ◽  
Rachel Lau ◽  
Filip Ralevski ◽  
Dylan R Pillai
Keyword(s):  
Quality Assurance ◽  
Real Time ◽  
Quantitative Pcr ◽  
Limit Of Detection ◽  
Performance Limit ◽  
Real Time Quantitative Pcr ◽  
Detection Analysis

scholarly journals Application of real-time quantitative PCR assays for detecting marine Brucella spp. in fish

2017 ◽  
Vol 30 (1) ◽  
pp. 150-154 ◽  
Author(s):  
Stephanie A. Norman ◽  
Martha A. Delaney ◽  
Katherine H. Haman ◽  
Austen C. Thomas ◽  
Jacques Godfroid ◽  
...  
Keyword(s):  
Real Time ◽  
Quantitative Pcr ◽  
Pacific Northwest ◽  
Marine Mammals ◽  
Atlantic Cod ◽  
Limit Of Detection ◽  
Hooded Seal ◽  
Real Time Quantitative Pcr ◽  
Zoonotic Infections ◽  
Paired Species

Brucella ceti and Brucella pinnipedialis have been documented as occurring in marine mammals, and B. ceti has been identified in 3 naturally acquired human cases. Seroconversion and infection patterns in Pacific Northwest harbor seals ( Phoca vitulina richardii) and North Atlantic hooded seals ( Cystophora cristata) indicate post-weaning exposure through prey consumption or lungworm infection, suggesting fish and possibly invertebrates play an epizootiologic role in marine Brucella transmission and possible foodborne risk to humans. We determined if real-time quantitative PCR (qPCR) assays can detect marine Brucella DNA in fish DNA. Insertion sequence (IS) 711 gene and sequence type (ST)27 primer–probe sets were used to detect Brucella associated with marine mammals and human zoonotic infections, respectively. First, DNA extracts from paired-species fish (containing 2 species) samples were tested and determined to be Brucella DNA negative using both IS 711 and ST27 primer–probe sets. A representative paired-species fish DNA sample was spiked with decreasing concentrations of B. pinnipedialis DNA to verify Brucella detection by the IS 711 primer–probe within fish DNA. A standard curve, developed using isolated DNA from B. pinnipedialis, determined the limit of detection. Finally, the IS 711 primer–probe was used to test Atlantic cod ( Gadus morhua) DNA extracts experimentally infected with the B. pinnipedialis hooded seal strain. In culture-positive cod tissue, the IS 711 limit of detection was ~1 genome copy of Brucella. Agreement between culture and PCR results for the 9 positive and 9 negative cod tissues was 100%. Although a larger sample set is required for validation, our study shows that qPCR can detect marine Brucella in fish.

scholarly journals Quantification and Stability of Human Adenoviruses and Polyomavirus JCPyV in Wastewater Matrices

2006 ◽  
Vol 72 (12) ◽  
pp. 7894-7896 ◽  
Author(s):  
Silvia Bofill-Mas ◽  
Nestor Albinana-Gimenez ◽  
Pilar Clemente-Casares ◽  
Ayalkibet Hundesa ◽  
Jesus Rodriguez-Manzano ◽  
...  
Keyword(s):  
Real Time ◽  
Quantitative Pcr ◽  
High Stability ◽  
Wastewater Sludge ◽  
Human Polyomavirus ◽  
Sewage Effluent ◽  
Viral Contamination ◽  
Human Adenoviruses ◽  
Urban Sewage ◽  
Real Time Quantitative Pcr

ABSTRACT Human adenoviruses (HAdV) and human polyomavirus JCPyV have been previously proposed as indicators of fecal viral contamination in the environment. Different wastewater matrices have been analyzed by applying real-time quantitative PCR procedures for the presence, quantity, and stability of a wide diversity of excreted HAdV and JCPyV. High quantities of HAdV and JCPyV were detected in sewage, effluent wastewater, sludge, and biosolid samples. Both viruses showed high stability in urban sewage. These results confirm the suitability of both viruses as indicators of human fecal viral pollution.

scholarly journals Identification, Detection, and Enumeration of Human Bifidobacterium Species by PCR Targeting the Transaldolase Gene

2002 ◽  
Vol 68 (5) ◽  
pp. 2420-2427 ◽  
Author(s):  
Teresa Requena ◽  
Jeremy Burton ◽  
Takahiro Matsuki ◽  
Karen Munro ◽  
Mary Alice Simon ◽  
...  
Keyword(s):  
Real Time ◽  
16S Rdna ◽  
Quantitative Pcr ◽  
Gene Sequence ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Bifidobacterium Longum ◽  
Bifidobacterium Adolescentis ◽  
Fecal Samples ◽  
Real Time Quantitative Pcr ◽  
Pcr Targeting

ABSTRACT Methods that enabled the identification, detection, and enumeration of Bifidobacterium species by PCR targeting the transaldolase gene were tested. Bifidobacterial species isolated from the feces of human adults and babies were identified by PCR amplification of a 301-bp transaldolase gene sequence and comparison of the relative migrations of the DNA fragments in denaturing gradient gel electrophoresis (DGGE). Two subtypes of Bifidobacterium longum, five subtypes of Bifidobacterium adolescentis, and two subtypes of Bifidobacterium pseudocatenulatum could be differentiated using PCR-DGGE. Bifidobacterium angulatum and B. catenulatum type cultures could not be differentiated from each other. Bifidobacterial species were also detected directly in fecal samples by this combination of PCR and DGGE. The number of species detected was less than that detected by PCR using species-specific primers targeting 16S ribosomal DNA (rDNA). Real-time quantitative PCR targeting a 110-bp transaldolase gene sequence was used to enumerate bifidobacteria in fecal samples. Real-time quantitative PCR measurements of bifidobacteria in fecal samples from adults correlated well with results obtained by culture when either a 16S rDNA sequence or the transaldolase gene sequence was targeted. In the case of samples from infants, 16S rDNA-targeted PCR was superior to PCR targeting the transaldolase gene for the quantification of bifidobacterial populations.

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