A rapid and high-throughput real-time PCR assay for species identification: application to stockfish sold in Italy

2009 ◽  
Vol 229 (2) ◽  
pp. 191-195 ◽  
Author(s):  
Gianluca Bertoja ◽  
Valerio Giaccone ◽  
Lisa Carraro ◽  
Alba N. Mininni ◽  
Barbara Cardazzo
Keyword(s):  
Real Time ◽  
Species Identification ◽  
High Throughput ◽  
Real Time Pcr ◽  
Pcr Assay

scholarly journals Development of a High-Throughput Quantitative Assay for Detecting Herpes Simplex Virus DNA in Clinical Samples

1999 ◽  
Vol 37 (6) ◽  
pp. 1941-1947 ◽  
Author(s):  
Alexander J. Ryncarz ◽  
James Goddard ◽  
Anna Wald ◽  
Meei-Li Huang ◽  
Bernard Roizman ◽  
...  
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Real Time ◽  
High Throughput ◽  
Real Time Pcr ◽  
Genital Tract ◽  
Clinical Samples ◽  
Pcr Assay ◽  
The Mean ◽  
Simplex Virus

We have developed a high-throughput, semiautomated, quantitative fluorescence-based PCR assay to detect and type herpes simplex virus (HSV) DNA in clinical samples. The detection assay, which uses primers to the type-common region of HSV glycoprotein B (gB), was linear from <10 to 108 copies of HSV DNA/20 μl of sample. Among duplicate samples in reproducibility runs, the assay showed less than 5% variability. We compared the fluorescence-based PCR assay with culture and gel-based liquid hybridization system with 335 genital tract specimens from HSV type 2 (HSV-2)-seropositive persons attending a research clinic and 380 consecutive cerebrospinal fluid (CSF) samples submitted to a diagnostic virology laboratory. Among the 162 culture-positive genital tract specimens, TaqMan PCR was positive for 157 (97%) specimens, whereas the quantitative-competitive PCR was positive for 144 (89%) specimens. Comparisons of the mean titer of HSV DNA detected by the two assays revealed that the mean titer detected by the gel-based system was slightly higher (median, 1 log). These differences in titers were in part related to the fivefold difference in the amount of HSV DNA used in the amplicon standards with the two assays. Among the 380 CSF samples, 42 were positive by both assays, 13 were positive only by the assay with the agarose gel, and 3 were positive only by the assay with the fluorescent probe. To define the subtype of HSV DNA detected in the screening assay, we also designed one set of primers which amplifies the gG regions of both types of HSV and probes which are specific to either HSV-1 (gG1) or HSV-2 (gG2). These probes were labeled with different fluorescent dyes (6-carboxyfluorescein for gG2 and 6-hexachlorofluorescein for gG1) to enable detection in a single PCR. In mixing experiments the probes discriminated the correct subtype in mixtures with up to a 7-log-higher concentration of the opposite subtype. The PCR typing results showed 100% concordance with the results obtained by assays with monoclonal antibodies against HSV-1 or HSV-2. Thus, while the real-time PCR is slightly less sensitive than the gel-based liquid hybridization system, the high throughput, the lack of contamination during processing, the better reproducibility, and the better ability to type the isolates rapidly make the real-time PCR a valuable tool for clinical investigation and diagnosis of HSV infection.

Species Identification of Fox-, Mink-, Dog-, and Rabbit-Derived Ingredients by Multiplex PCR and Real-Time PCR Assay

2017 ◽  
Vol 185 (1) ◽  
pp. 1-12 ◽  
Author(s):  
Qingqing Wu ◽  
Shengnan Xiang ◽  
Wenjun Wang ◽  
Jinyan Zhao ◽  
Jinhua Xia ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Real Time ◽  
Species Identification ◽  
Real Time Pcr ◽  
Pcr Assay

High-throughput two-step LNA real time PCR assay for the quantitative detection and genotyping of HPV prognostic-risk groups

2009 ◽  
Vol 45 (4) ◽  
pp. 304-310 ◽  
Author(s):  
Elisa Leo ◽  
Simona Venturoli ◽  
Monica Cricca ◽  
Monica Musiani ◽  
Marialuisa Zerbini
Keyword(s):  
Real Time ◽  
High Throughput ◽  
Real Time Pcr ◽  
Risk Groups ◽  
Quantitative Detection ◽  
Pcr Assay

scholarly journals Detection and species identification of microsporidial infections using SYBR Green real-time PCR

2011 ◽  
Vol 60 (4) ◽  
pp. 459-466 ◽  
Author(s):  
Spencer D. Polley ◽  
Samuel Boadi ◽  
Julie Watson ◽  
Alan Curry ◽  
Peter L. Chiodini
Keyword(s):  
Real Time ◽  
Species Identification ◽  
Real Time Pcr ◽  
Simultaneous Detection ◽  
Enterocytozoon Bieneusi ◽  
Sybr Green ◽  
Pcr Assay ◽  
Real Time Analysis ◽  
Highly Sensitive ◽  
Stool Samples

Diagnosis of microsporidial infections is routinely performed by light microscopy, with unequivocal non-molecular species identification achievable only through electron microscopy. This study describes a single SYBR Green real-time PCR assay for the simultaneous detection and species identification of such infections. This assay was highly sensitive, routinely detecting infections containing 400 parasites (g stool sample)−1, whilst species identification was achieved by differential melt curves on a Corbett Life Science Rotor-Gene 3000. A modification of the QIAamp DNA tissue extraction protocol allowed the semi-automated extraction of DNA from stools for the routine diagnosis of microsporidial infection by real-time PCR. Of 168 stool samples routinely analysed for microsporidian spores, only five were positive by microscopy. By comparison, 17 were positive for microsporidial DNA by real-time analysis, comprising 14 Enterocytozoon bieneusi, one Encephalitozoon cuniculi and two separate Pleistophora species infections.

A fluorescence-based quantitative real-time PCR assay for accurate Pocillopora damicornis species identification

Coral Reefs ◽  
2016 ◽  
Vol 35 (3) ◽  
pp. 895-899 ◽  
Author(s):  
Luke Thomas ◽  
Michael Stat ◽  
Richard D. Evans ◽  
W. Jason Kennington
Keyword(s):  
Real Time ◽  
Species Identification ◽  
Real Time Pcr ◽  
Pocillopora Damicornis ◽  
Quantitative Real Time Pcr ◽  
Pcr Assay

A high-throughput microfluidic biochip to quantify bacterial adhesion to single host cells by real-time PCR assay

2013 ◽  
Vol 405 (12) ◽  
pp. 4277-4282 ◽  
Author(s):  
Rui Zhang ◽  
Hai-Qing Gong ◽  
Xu Dong Zeng ◽  
Chun Chau Sze
Keyword(s):  
Real Time ◽  
High Throughput ◽  
Real Time Pcr ◽  
Bacterial Adhesion ◽  
Host Cells ◽  
Pcr Assay ◽  
Single Host

scholarly journals High-throughput multiplex real-time PCR assay for the simultaneous quantification of DNA and RNA viruses infecting cassava plants

2016 ◽  
Vol 120 (5) ◽  
pp. 1346-1356 ◽  
Author(s):  
G. Otti ◽  
S. Bouvaine ◽  
B. Kimata ◽  
G. Mkamillo ◽  
P.L. Kumar ◽  
...  
Keyword(s):  
Real Time ◽  
High Throughput ◽  
Real Time Pcr ◽  
Rna Viruses ◽  
Pcr Assay ◽  
Simultaneous Quantification ◽  
Dna And Rna

Efficient species identification of pine marten (Martes martes) and red fox (Vulpes vulpes) scats using a 5′ nuclease real-time PCR assay

2007 ◽  
Vol 9 (3) ◽  
pp. 735-738 ◽  
Author(s):  
Catherine O’Reilly ◽  
Mark Statham ◽  
Jacinta Mullins ◽  
Peter D. Turner ◽  
Declan O’Mahony
Keyword(s):  
Real Time ◽  
Species Identification ◽  
Real Time Pcr ◽  
Vulpes Vulpes ◽  
Red Fox ◽  
Martes Martes ◽  
Pine Marten ◽  
Pcr Assay

scholarly journals qPCRTag Analysis - A High Throughput, Real Time PCR Assay for Sc2.0 Genotyping

10.3791/52941 ◽  
2015 ◽  
Author(s):  
Leslie A. Mitchell ◽  
Nick A. Phillips ◽  
Andrea Lafont ◽  
James A. Martin ◽  
Rupal Cutting ◽  
...  
Keyword(s):  
Real Time ◽  
High Throughput ◽  
Real Time Pcr ◽  
Pcr Assay
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