scholarly journals Diagnostic Real-Time PCR Assays for the Detection of Emetic Bacillus cereus Strains in Foods and Recent Food-Borne Outbreaks

2007 ◽  
Vol 73 (6) ◽  
pp. 1892-1898 ◽  
Author(s):  
Martina Fricker ◽  
Ute Messelhäußer ◽  
Ulrich Busch ◽  
Siegfried Scherer ◽  
Monika Ehling-Schulz
Keyword(s):  
Bacillus Cereus ◽  
Real Time ◽  
Real Time Pcr ◽  
Care Center ◽  
Food Poisoning ◽  
Sybr Green ◽  
Sybr Green I ◽  
Taqman Assay ◽  
Pcr Assay ◽  
Food Borne

ABSTRACT Cereulide-producing Bacillus cereus can cause an emetic type of food-borne disease that mimics the symptoms provoked by Staphylococcus aureus. Based on the recently discovered genetic background for cereulide formation, a novel 5′ nuclease (TaqMan) real-time PCR assay was developed to provide a rapid and sensitive method for the specific detection of emetic B. cereus in food. The TaqMan assay includes an internal amplification control and primers and a probe designed to target a highly specific part of the cereulide synthetase genes. Additionally, a specific SYBR green I assay was developed and extended to create a duplex SYBR green I assay for the one-step identification and discrimination of the two emesis-causing food pathogens B. cereus and S. aureus. The inclusivity and exclusivity of the assay were assessed using a panel of 100 strains, including 23 emetic B. cereus and 14 S. aureus strains. Different methods for DNA isolation from artificially contaminated foods were evaluated, and established real-time assays were used to analyze two recent emetic food poisonings in southern Germany. One of the food-borne outbreaks included 17 children visiting a day care center who vomited after consuming a reheated rice dish, collapsed, and were hospitalized; the other case concerned a single food-poisoning incident occurring after consumption of cauliflower. Within 2 h, the etiological agent of these food poisonings was identified as emetic B. cereus by using the real-time PCR assay.

scholarly journals Development of a SYBR green I-based duplex real-time PCR assay for detection of pseudorabies virus and porcine circovirus 3

2020 ◽  
Vol 53 ◽  
pp. 101593 ◽  
Author(s):  
Run-Bo Tian ◽  
Yue Jin ◽  
Tong Xu ◽  
Yu Zhao ◽  
Zhen-Ya Wang ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Pseudorabies Virus ◽  
Sybr Green ◽  
Sybr Green I ◽  
Porcine Circovirus ◽  
Pcr Assay ◽  
Porcine Circovirus 3

Fast and sensitive quantitative detection of HIV DNA in whole blood leucocytes by SYBR green I real-time PCR assay

2007 ◽  
Vol 21 (5-6) ◽  
pp. 368-378 ◽  
Author(s):  
Anna Casabianca ◽  
Caterina Gori ◽  
Chiara Orlandi ◽  
Federica Forbici ◽  
Carlo Federico Perno ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Whole Blood ◽  
Sybr Green ◽  
Sybr Green I ◽  
Quantitative Detection ◽  
Pcr Assay ◽  
Hiv Dna

scholarly journals Quantitative Detection of Listeria monocytogenes in Biofilms by Real-Time PCR

2005 ◽  
Vol 71 (4) ◽  
pp. 2190-2194 ◽  
Author(s):  
Morgan Guilbaud ◽  
Pierre de Coppet ◽  
Fabrice Bourion ◽  
Cinta Rachman ◽  
Hervé Prévost ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Real Time ◽  
Real Time Pcr ◽  
Quantitative Method ◽  
Sybr Green ◽  
Sybr Green I ◽  
Quantitative Detection ◽  
Pcr Assay

ABSTRACT A quantitative method based on a real-time PCR assay to enumerate Listeria monocytogenes in biofilms was developed. The specificity for L. monocytogenes of primers targeting the listeriolysin gene was demonstrated using a SYBR Green I real-time PCR assay. The number of L. monocytogenes detected growing in biofilms was 6 × 102 CFU/cm2.

Development of a SYBR Green I based one-step real-time PCR assay for the detection of Hantaan virus

2014 ◽  
Vol 196 ◽  
pp. 145-151 ◽  
Author(s):  
Wei Jiang ◽  
Ping-zhong Wang ◽  
Hai-tao Yu ◽  
Ye Zhang ◽  
Ke Zhao ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Sybr Green ◽  
Sybr Green I ◽  
Hantaan Virus ◽  
Pcr Assay ◽  
One Step

Real-time PCR assay for sensitive organ detection and epidemic investigation of Turbot reddish body iridovirus

2009 ◽  
Vol 6 (1) ◽  
pp. 61-67 ◽  
Author(s):  
Wu Cheng-Long ◽  
Shi Cheng-Yin ◽  
Huang Jie ◽  
Kong Xiao-Yu
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Viral Genome ◽  
Sybr Green ◽  
Sybr Green I ◽  
Quantitative Detection ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Linear Coefficient ◽  
Sensitive Organ

AbstractA rapid and sensitive real-time polymerase chain reaction (PCR) assay coupled with SYBR Green I chemistry was developed for the quantitative detection of Turbot reddish body iridovirus (TRBIV) isolated from farmed turbot (Scophthalmus maximus). A 152 bp DNA fragment from the TRBIV major capsid protein (MCP) gene was involved in the real-time PCR (RT-PCR) assay using the Roter Gene 3000 sequence detection system. The PCR mixture contained a fluorescent dye, SYBR Green I, which exhibited fluorescence enhancement when bound to double-stranded (ds) DNA. The enhancement of fluorescence was proportional to the initial concentration of the template DNA. The positive control plasmid, pUCm-T/TRBIV MCP, containing the target sequence, was quantified to make a standard curve for sample detection after serial tenfold dilution. Linear coefficient correlations between the cycle threshold (CT) value and logarithmic positive plasmid concentration were close to one (R2=0.9952) and the detection limit of the assay was 102 copies of positive plasmids. The quantitative detection of virus in different tissues from TRBIV-infected fish showed that the spleen and kidney contained the largest number of viral particles (5.23×106 and 2.18×106 viral genome copies/mg tissue, respectively), while no viral DNA was detected in the muscular tissue. The molecular epidemic investigation of TRBIV showed that many cultured turbots were infected and TRBIV has become epidemic in turbot farms located along the Shandong peninsula. The virus number varied from 1.27×102 to 2.33×106 viral genome copies/mg tissue in spleens of infected turbot. These results suggest that the RT-PCR assay reported here can be used as a rapid, sensitive and quantitative method for TRBIV.

Development of an SYBR Green I real-time PCR assay for detection ofEdwardsiella tardaand its application

2013 ◽  
Vol 37 (12) ◽  
pp. 1829 ◽  
Author(s):  
Xiaojun RONG ◽  
Meijie LIAO ◽  
Zheng ZHANG ◽  
Yingeng WANG ◽  
Zhichao LIU ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Sybr Green ◽  
Sybr Green I ◽  
Pcr Assay

scholarly journals Development of an SYBR Green I-based real-time PCR assay for the rapid detection of canine kobuvirus

2020 ◽  
Vol 285 ◽  
pp. 113944
Author(s):  
Yong Wang ◽  
Yongqiu Cui ◽  
Yeqiu Li ◽  
Da Zhang ◽  
Jianfei Sun ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Rapid Detection ◽  
Sybr Green ◽  
Sybr Green I ◽  
Pcr Assay ◽  
Canine Kobuvirus
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