Molecular characterization of toxigenic clostridium difficile by multiplex and real-time PCR

2011 ◽  
Author(s):  
Chi-chiu, Elvin Chan
Keyword(s):  
Clostridium Difficile ◽  
Real Time ◽  
Molecular Characterization ◽  
Real Time Pcr

scholarly journals Molecular Characterization of Giardia duodenalis in Children and Adults Sampled in Algeria

2020 ◽  
Vol 9 (1) ◽  
pp. 54
Author(s):  
Salem Belkessa ◽  
Daniel Thomas-Lopez ◽  
Karim Houali ◽  
Farida Ghalmi ◽  
Christen Rune Stensvold
Keyword(s):  
Real Time ◽  
Molecular Characterization ◽  
Real Time Pcr ◽  
Intestinal Parasites ◽  
Giardia Duodenalis ◽  
Specific Pcr ◽  
Stool Samples ◽  
Pcr Targeting ◽  
Assemblage B

The molecular epidemiology of giardiasis in Africa remains unclear. A study was carried out across four hospitals in Algeria. A total of 119 fecal samples from 55 children, 37 adults, and 27 individuals of undetermined age, all scored positive for intestinal parasites by microscopy, and were screened by real-time PCR for Giardia. Molecular characterization of Giardia was performed by assemblage-specific PCR and PCR targeting the triose phosphate isomerase gene (tpi). Of the 119 samples, 80 (67%) were Giardia-positive by real-time PCR. For 48 moderately-highly real-time PCR-positive samples, tpi genotyping assigned 22 samples to Assemblage A and 26 to Assemblage B. Contrary to Assemblage A, Assemblage B exhibited substantial genetic diversity and allelic heterozygosity. Assemblage-specific PCR proved to be specific for discriminating Assemblage A or B but not as sensitive as tpi genotyping. We confirmed that real-time PCR is more sensitive than microscopy for detecting Giardia in stool samples and that robust amplification and sequencing of the tpi gene is feasible when moderate-to-strongly real-time PCR-positive samples are used. This study is one of the few performed in Africa providing genotyping data on Giardia infections in humans. Both assemblages A and B were commonly seen and not associated with specific sociodemographic data.

scholarly journals Multiplex Real-Time PCR Method for Simultaneous Identification and Toxigenic Type Characterization of Clostridium difficile From Stool Samples

2015 ◽  
Vol 35 (3) ◽  
pp. 306-313 ◽  
Author(s):  
Abdullah Kilic ◽  
Mohammad J. Alam ◽  
Naradah L. Tisdel ◽  
Dhara N. Shah ◽  
Mehmet Yapar ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Real Time ◽  
Real Time Pcr ◽  
Stool Samples ◽  
Pcr Method ◽  
Simultaneous Identification

Molecular Characterization of Prader–Willi Syndrome by Real-Time PCR

2008 ◽  
Vol 12 (2) ◽  
pp. 319-324 ◽  
Author(s):  
Teresa Munce ◽  
Robert Simpson ◽  
Francis Bowling
Keyword(s):  
Real Time ◽  
Molecular Characterization ◽  
Real Time Pcr ◽  
Prader Willi Syndrome

scholarly journals Advances in molecular surveillance of Clostridium difficile in Bulgaria

2013 ◽  
Vol 62 (9) ◽  
pp. 1428-1434 ◽  
Author(s):  
Elina G. Dobreva ◽  
Ivan N. Ivanov ◽  
Rossitza S. Vathcheva-Dobrevska ◽  
Katucha I. Ivanova ◽  
Galina D. Asseva ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Real Time ◽  
Real Time Pcr ◽  
Simultaneous Detection ◽  
Variable Number ◽  
Innovative Methods ◽  
Pcr Ribotyping ◽  
Typing Methods ◽  
Stool Samples

The increasing incidence of Clostridium difficile infection (CDI) in Bulgaria has indicated the need to implement better surveillance approaches. The aim of the present work was to improve the current surveillance of CDI in Bulgaria by introducing innovative methods for identification and typing. One hundred and twenty stool samples obtained from 108 patients were studied over 4 years from which 32 C. difficile isolates were obtained. An innovative duplex EvaGreen real-time PCR assay based on simultaneous detection of the gluD and tcdB genes was developed for rapid C. difficile identification. Four toxigenic profiles were distinguished by PCR: A+B+CDT− (53.1 %, 17/32), A−B+CDT− (28.1 %, 9/32), A+B+CDT+ (9.4 %, 3/32) and A−B−CDT− (9.4 %, 3/32). PCR ribotyping and multilocus variable number of tandem repeat analysis (MLVA7) were used for molecular characterization of the isolates. In total, nine distinct ribotypes were confirmed and the most prevalent for Bulgarian hospitals was 017 followed by 014/020, together accounting for 44 % of all isolates. Eighteen per cent of the isolates (6/32) did not match any of the 25 reference ribotypes available in this study. Twenty-four MLVA7 genotypes were detected among the clinical C. difficile isolates, distributed as follows: five for 017 ribotype, two for 014/020, 001, 002, 012 and 046 each, and one each for ribotypes 023, 070 and 078. The correlation between the typing methods was significant and allowed the identification of several clonal complexes. These results suggest that most C. difficile cases in the eight Bulgarian hospitals studied were associated with isolates belonging to the outbreak ribotypes 017 and 014/20, which are widely distributed in Europe. The real-time PCR protocol for simultaneous detection of gluD and tcdB proved to be very effective and improved C. difficile identification and confirmation of clinical C. difficile isolates.

scholarly journals Molecular characterization of circulating tumor cells in large quantities of contaminating leukocytes by a multiplex real-time PCR

2008 ◽  
Vol 118 (3) ◽  
pp. 455-468 ◽  
Author(s):  
Anieta M. Sieuwerts ◽  
Jaco Kraan ◽  
Joan Bolt-de Vries ◽  
Petra van der Spoel ◽  
Bianca Mostert ◽  
...  
Keyword(s):  
Real Time ◽  
Tumor Cells ◽  
Molecular Characterization ◽  
Circulating Tumor Cells ◽  
Real Time Pcr

Molecular Characterization of Large Deletions in the von Hippel-Lindau (VHL) Gene by Quantitative Real-Time PCR

2006 ◽  
Vol 10 (4) ◽  
pp. 243-249 ◽  
Author(s):  
Alberto Casarin ◽  
Maddalena Martella ◽  
Roberta Polli ◽  
Emanuela Leonardi ◽  
Laura Anesi ◽  
...  
Keyword(s):  
Real Time ◽  
Molecular Characterization ◽  
Real Time Pcr ◽  
Quantitative Real Time Pcr ◽  
Vhl Gene ◽  
Von Hippel Lindau ◽  
Large Deletions

Molecular characterization of atypical Chlamydia and evidence of their dissemination in different European and Asian chicken flocks by specific real-time PCR

2012 ◽  
Vol 14 (8) ◽  
pp. 2212-2222 ◽  
Author(s):  
Aleksandar Zocevic ◽  
Fabien Vorimore ◽  
Cvetka Marhold ◽  
Danijela Horvatek ◽  
Dongying Wang ◽  
...  
Keyword(s):  
Real Time ◽  
Molecular Characterization ◽  
Real Time Pcr

Real-time PCR-based prevalence study, infection follow-up and molecular characterization of canine hemotropic mycoplasmas

2008 ◽  
Vol 126 (1-3) ◽  
pp. 132-141 ◽  
Author(s):  
Nicole Wengi ◽  
Barbara Willi ◽  
Felicitas S. Boretti ◽  
Valentino Cattori ◽  
Barbara Riond ◽  
...  
Keyword(s):  
Real Time ◽  
Molecular Characterization ◽  
Real Time Pcr ◽  
Prevalence Study
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