TaqMan minor groove binder real-time PCR analysis of β-tubulin codon 200 polymorphism in small strongyles (Cyathostomin) indicates that the TAC allele is only moderately selected in benzimidazole-resistant populations

Parasitology ◽  
2003 ◽  
Vol 127 (5) ◽  
pp. 489-496 ◽  
Author(s):  
G. VON SAMSON-HIMMELSTJERNA ◽  
S. BUSCHBAUM ◽  
N. WIRTHERLE ◽  
M. PAPE ◽  
T. SCHNIEDER
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Minor Groove ◽  
Pcr Analysis ◽  
Minor Groove Binder ◽  
Allelic Discrimination ◽  
The Real ◽  
Larval Stages ◽  
Taqman Minor Groove Binder ◽  
Β Tubulin

TaqMan minor groove binder probes were evaluated as to their suitability for the real-time allelic discrimination of the β-tubulin codon 200 TTC/TAC single nucleotide polymorphism in cyathostomin species. Amplification of titrated cloned full-length β-tubulin cDNA revealed that the TaqMan minor groove binder PCR is capable of specifically detecting as few as 10 copies. Testing of DNA from single adult and larval stages of several different species of cyathostomin allowed reproducible genotyping of individual worms. Using the real-time PCR approach, the throughput of samples was considerably increased compared with conventional post-PCR readout procedure. Only 7·8% homozygous TAC L3 were found among 102 L3 which were genotyped from phenotypically BZ-resistant small strongyle populations. The percentages of the homozygous TTC and heterozygous TTC/TAC were 41·3% and 50·9%, respectively. This resulted in a total TAC-allele percentage of only 33·3%. These findings correspond to data obtained by genotyping of an experimentally selected BZ-resistant cyathostomin population. It is concluded that the β-tubulin codon 200 polymorphism is not the sole mechanism involved in the process of BZ resistance in cyathostomins.

scholarly journals Touchable 3D hierarchically structured polyaniline nanoweb for capture and detection of pathogenic bacteria

2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Kyung Hoon Kim ◽  
MinHo Yang ◽  
Younseong Song ◽  
Chi Hyun Kim ◽  
Young Mee Jung ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Pathogenic Bacteria ◽  
Bacterial Pathogens ◽  
Physical Contact ◽  
Structural Deformation ◽  
Pcr Analysis ◽  
Mechanical Resistance ◽  
Fluorescence Signal ◽  
The Real

AbstractA bacteria-capturing platform is a critical function of accurate, quantitative, and sensitive identification of bacterial pathogens for potential usage in the detection of foodborne diseases. Despite the development of various nanostructures and their surface chemical modification strategies, relative to the principal physical contact propagation of bacterial infections, mechanically robust and nanostructured platforms that are available to capture bacteria remain a significant problem. Here, a three-dimensional (3D) hierarchically structured polyaniline nanoweb film is developed for the efficient capture of bacterial pathogens by hand-touching. This unique nanostructure ensures sufficient mechanical resistance when exposed to compression and shear forces and facilitates the 3D interfacial interactions between bacterial extracellular organelles and polyaniline surfaces. The bacterial pathogens (Escherichia coli O157:H7, Salmonella enteritidis, and Staphylococcus aureus) are efficiently captured through finger-touching, as verified by the polymerase chain reaction (PCR) analysis. Moreover, the real-time PCR results of finger-touched cells on a 3D nanoweb film show a highly sensitive detection of bacteria, which is similar to those of the real-time PCR using cultured cells without the capturing step without any interfering of fluorescence signal and structural deformation during thermal cycling. Graphic Abstract

scholarly journals Detection of Elm Yellows Phytoplasma in Elms and Insects Using Real-Time PCR

Plant Disease ◽  
2010 ◽  
Vol 94 (11) ◽  
pp. 1355-1360 ◽  
Author(s):  
Padmini Herath ◽  
Gregory A. Hoover ◽  
Elisa Angelini ◽  
Gary W. Moorman
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Extraction Procedure ◽  
Accurate Method ◽  
The United States ◽  
State University ◽  
Phylogenetic Groups ◽  
The Real ◽  
Elm Yellows ◽  
Taqman Minor Groove Binder

A rapid and accurate method to detect the common strain of elm yellows (EY) phytoplasma in elm and insect samples was developed using a real-time polymerase chain reaction (PCR) procedure based on the TaqMan minor-groove-binder probe. Primers and probe were designed based on the EY phytoplasma-specific translocation protein secY gene DNA sequence. Success of the DNA extraction procedure was evaluated by amplifying the chloroplast trnL gene of Ulmus americana. The real-time PCR assay reacted positively with EY and EY phytoplasma strain ULW DNA, an isolate which occurs in Europe. It did not cross-react with Illinois EY or aster yellows phytoplasma DNA, both of which are known to occur in elm trees in the United States, nor did it amplify several other phytoplasmas belonging to the 16SrV and other phylogenetic groups. The real-time PCR protocol was used to identify 30 EY-positive elm trees on The Pennsylvania State University, University Park campus. Threshold cycle (CT) values obtained from the EY phytoplasma-infected elm trees ranged from 15 to 37. EY phytoplasma was detected in several leafhopper taxa. This real-time PCR method can be used for the diagnostic screening of elm trees and for the identification of possible insect vectors of EY phytoplasma.

scholarly journals A minor groove binder probe real-time PCR assay for discrimination between type 2-based vaccines and field strains of canine parvovirus

2006 ◽  
Vol 136 (1-2) ◽  
pp. 65-70 ◽  
Author(s):  
Nicola Decaro ◽  
Gabriella Elia ◽  
Costantina Desario ◽  
Sante Roperto ◽  
Vito Martella ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Minor Groove ◽  
Canine Parvovirus ◽  
Minor Groove Binder ◽  
Pcr Assay ◽  
Minor Groove Binder Probe ◽  
A Minor
Sign in / Sign up

Export Citation Format

Share Document