Enhanced enrichment and detection of thermotolerant Campylobacter species from water using the Portable Microbe Enrichment Unit and real-time PCR

2009 ◽  
Vol 55 (7) ◽  
pp. 849-858 ◽  
Author(s):  
Tarja Pitkänen ◽  
Juliane Bräcker ◽  
Ilkka T. Miettinen ◽  
Anneli Heitto ◽  
Jouni Pesola ◽  
...  
Keyword(s):  
Drinking Water ◽  
Real Time ◽  
Campylobacter Jejuni ◽  
Real Time Pcr ◽  
Pcr Detection ◽  
Detection Time ◽  
The Real ◽  
International Standard ◽  
Campylobacter Species

An enhanced enrichment using the Portable Microbe Enrichment Unit (PMEU) with the microaerobic bubbling of broths was applied for the detection of thermotolerant Campylobacter species from water. This PMEU enrichment was compared with the conventional static enrichment of the international standard ISO 17995:2005. In addition, Campylobacter detection after enrichment using a real-time PCR detection was compared with colony counts. The tests with stressed Campylobacter jejuni cells in drinking water indicated that the PMEU enrichment yielded a significantly higher number of Campylobacter cells in the Bolton broth compared with the conventional static incubation. Application of the real-time PCR technique shortened the Campylobacter detection time. This combination of method modifications can be used for Campylobacter detection from water and adds methodological repertoire for the rapid survey and management of waterborne outbreaks.

Combined effects of matrix and gene marker on the real-time PCR detection of wheat

2016 ◽  
Vol 51 (7) ◽  
pp. 1680-1688 ◽  
Author(s):  
Begoña Martín-Fernández ◽  
Joana Costa ◽  
Maria Beatriz Prior Pinto Oliveira ◽  
Beatriz López-Ruiz ◽  
Isabel Mafra
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Pcr Detection ◽  
Gene Marker ◽  
Combined Effects ◽  
The Real

Quantification and Differentiation of Campylobacter jejuni and Campylobacter coli in Raw Chicken Meats Using a Real-Time PCR Method

2007 ◽  
Vol 70 (9) ◽  
pp. 2015-2022 ◽  
Author(s):  
JOONBAE HONG ◽  
WOO KYUNG JUNG ◽  
JUN MAN KIM ◽  
SO HYUN KIM ◽  
HYE CHEONG KOO ◽  
...  
Keyword(s):  
Real Time ◽  
Quantitative Pcr ◽  
Real Time Pcr ◽  
Simultaneous Detection ◽  
High Specificity ◽  
Campylobacter Coli ◽  
Pcr Assay ◽  
The Real ◽  
Real Time Quantitative Pcr ◽  
Campylobacter Species

Campylobacter species are one of the most common causes of bacterial diarrhea in humans worldwide. The consumption of foods contaminated with two Campylobacter species, C. jejuni and C. coli, is usually associated with most of the infections in humans. In this study, a rapid, reliable, and sensitive multiplex real-time quantitative PCR was developed for the simultaneous detection, identification, and quantification of C. jejuni and C. coli. In addition, the developed method was applied to the 50 samples of raw chicken meat collected from retail stores in Korea. C. jejuni and C. coli were detected in 88 and 86% of the samples by real-time quantitative PCR and the conventional microbiological method, respectively. The specificity of the primer and probe sets was confirmed with 30 C. jejuni, 20 C. coli, and 35 strains of other microbial species. C. jejuni and C. coli could be detected with high specificity in less than 4 h, with a detection limit of 1 log CFU/ml by the developed real-time PCR. The average counts (log CFU per milliliter) of C. jejuni or C. coli obtained by the conventional methods and by the real-time PCR assay were statistically correlated with a correlation coefficient (R2) between 0.73 and 0.78. The real-time PCR assay developed in this study is useful for screening for the presence and simultaneous differential quantification of C. jejuni and C. coli.

Real-Time PCR Detection of Enteric Viruses in Source Water and Treated Drinking Water in Wuhan, China

2012 ◽  
Vol 65 (3) ◽  
pp. 244-253 ◽  
Author(s):  
Xiao Yan Ye ◽  
Xing Ming ◽  
Yong Lu Zhang ◽  
Wen Qing Xiao ◽  
Xia Ning Huang ◽  
...  
Keyword(s):  
Drinking Water ◽  
Real Time ◽  
Real Time Pcr ◽  
Enteric Viruses ◽  
Pcr Detection ◽  
Source Water ◽  
Treated Drinking Water

scholarly journals Rectal Swabs as an Alternative Sample Collection Method to Bulk Stool for the Real-Time PCR Detection of Giardia duodenalis

2020 ◽  
Vol 103 (3) ◽  
pp. 1276-1282 ◽  
Author(s):  
Jacqueline R. M. A. Maasch ◽  
Ahmed M. Arzika ◽  
Catherine Cook ◽  
Elodie Lebas ◽  
Nils Pilotte ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Giardia Duodenalis ◽  
Pcr Detection ◽  
Sample Collection ◽  
Collection Method ◽  
The Real ◽  
Rectal Swabs

scholarly journals A Real-Time PCR Assay for the Detection of Campylobacter jejuni in Foods after Enrichment Culture

2003 ◽  
Vol 69 (3) ◽  
pp. 1383-1390 ◽  
Author(s):  
Andrew D. Sails ◽  
Andrew J. Fox ◽  
Frederick J. Bolton ◽  
David R. A. Wareing ◽  
David L. A. Greenway
Keyword(s):  
Real Time ◽  
Campylobacter Jejuni ◽  
Real Time Pcr ◽  
Enrichment Culture ◽  
Poultry Meat ◽  
Pcr Assay ◽  
Enrichment Broth ◽  
The Real ◽  
Enrichment Cultures ◽  
Selective Agar

ABSTRACT A real-time PCR assay was developed for the quantitative detection of Campylobacter jejuni in foods after enrichment culture. The specificity of the assay for C. jejuni was demonstrated with a diverse range of Campylobacter species, related organisms, and unrelated genera. The assay had a linear range of quantification over six orders of magnitude, and the limit of detection was approximately 12 genome equivalents. The assay was used to detect C. jejuni in both naturally and artificially contaminated food samples. Ninety-seven foods, including raw poultry meat, offal, raw shellfish, and milk samples, were enriched in blood-free Campylobacter enrichment broth at 37°C for 24 h, followed by 42°C for 24 h. Enrichment cultures were subcultured to Campylobacter charcoal-cefoperazone-deoxycholate blood-free selective agar, and presumptive Campylobacter isolates were identified with phenotypic methods. DNA was extracted from enrichment cultures with a rapid lysis method and used as the template in the real-time PCR assay. A total of 66 samples were positive for C. jejuni by either method, with 57 samples positive for C. jejuni by subculture to selective agar medium and 63 samples positive in the real-time PCR assay. The results of both methods were concordant for 84 of the samples. The total time taken for detection from enrichment broth samples was approximately 3 h for the real-time PCR assay, with the results being available immediately at the end of PCR cycling, compared to 48 h for subculture to selective agar. This assay significantly reduces the total time taken for the detection of C. jejuni in foods and is an important model for other food-borne pathogens.

The Establishment of Real-Time PCR Detection System of Aspergillus Niger in Peanut

2016 ◽  
Vol 835 ◽  
pp. 333-337
Author(s):  
Chu Shu Zhang ◽  
Jie Sun ◽  
Li Na Yu ◽  
Jie Bi ◽  
Wei Wei Zhao
Keyword(s):  
Aspergillus Niger ◽  
Real Time ◽  
Correlation Coefficient ◽  
Real Time Pcr ◽  
Detection System ◽  
Dna Detection ◽  
Pcr Primers ◽  
Pcr Detection ◽  
The Real

In this study, a set of real-time PCR primers for the detection of aflatoxin in peanut was successfully designed according to the AFLR gene of Aspergillus niger. Upstream primer was 5'-AACCTGATGACGACTGATAT-3'; Downstream primer was 5'-AGCACCTTGAGAACGATAA-3'; The real-time PCR detection system established in this study had good specificity, and the sensitivity of DNA detection can reach 3×10-13 ng/ul. The correlation coefficient between the contents of aflatoxin in peanut samples and PCR Ct of Real-time was-0.718, and the correlation coefficient was significantly negative.

APPLICATION OF REAL-TIME PRC TECHNIQUE AND REVERSE DOT-BLOT FOR DETECTION THE HIGH RISK TYPES OF HUMAN PAPILLOMAVIRUS CAUSING CERVICAL CANCER

2017 ◽  
pp. 99-103
Author(s):  
Van Bao Thang Phan ◽  
Hoang Bach Nguyen ◽  
Van Thanh Nguyen ◽  
Thi Nhu Hoa Tran ◽  
Viet Quynh Tram Ngo
Keyword(s):  
Cervical Cancer ◽  
High Risk ◽  
Real Time ◽  
Real Time Pcr ◽  
Dot Blot ◽  
The Real ◽  
Dot Blot Assay ◽  
Hpv Types ◽  
High Risk Hpv ◽  
Reverse Dot Blot

Introduction: Infection with HPV is the main cause of cervical cancer. Determining HPV infection and the types of HPV plays an important role in diagnosis, treatment and prognosis of cervicitis/cervical cancer. Aims: Determining proportion of high-risk HPV types and the occurrence of coinfection with multiple HPV types. Methods: 177 women with cervicitis or abnormal Pap smear result were enrolled in the study. Performing the real-time PCR for detecting HPV and the reverse DOT-BLOT assay for determining type of HPV in cases of positive PCR. Results: 7 types of high-risk HPV was dectected, the majority of these types were HPV type 18 (74.6%) and HPV type 16 (37.6%); the proportion of infection with only one type of HPV was 30.4% and coinfection with multiple HPV types was higher (69.6%), the coinfected cases with 2 and 3 types were dominated (32.2% and 20.3%, respectively) and the coinfected cases with 4 and 5 types were rare. Conclusion: Use of the real-time PCR and reverse DOT-BLOT assay can determine the high-risk HPV types and the occurrence of coinfection with multiple HPV types. Key words: HPV type, Reverse DOT-BLOT, real-time PCR,PCR, cervical cancer

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