scholarly journals Monitoring Living Modified Canola Using an Efficient Multiplex PCR Assay in Natural Environments in South Korea

2020 ◽  
Vol 10 (21) ◽  
pp. 7721
Author(s):  
Il Ryong Kim ◽  
Hye Song Lim ◽  
Wonkyun Choi ◽  
Da In Kang ◽  
Sang Yeol Lee ◽  
...  
Keyword(s):  
South Korea ◽  
Multiplex Pcr ◽  
Natural Environments ◽  
Pcr Assay ◽  
Brassica Napus L ◽  
Livestock Feed ◽  
Multiplex Pcr Assay ◽  
Management Project ◽  
Pcr Method ◽  
Living Modified Organisms

Canola (Brassica napus L.) is cultivated worldwide and utilized as a vegetable oil, biodiesel, and livestock feed. It is also a major living modified (LM) crop alongside corn, soybean, and cotton. Many canola events have been authorized for food, feed, and processing use in South Korea. Concerns about the unintentional release of LM canola into the natural environment have increased environmental monitoring and post-management of living modified organisms (LMOs) is on the rise. The Ministry of Environment (MOE) and the National Institute of Ecology (NIE) conducted an environmental LMO monitoring and post-management project for LM canola from 2014 to 2017. The number of suspicious LM samples gradually increased each year. In this study, a multiplex PCR method was established to detect seven single LM canola events (Topas 19/2, Rf3, Dp-73496-4, Ms8, GT73, Mon88032, and T45) to cover 14 approved LM canola events. This method was utilized to detect 22 LMs out of 260 suspicious canola samples. Thus, this new method is more efficient in terms of time and cost than conventional PCR methods for the identification and monitoring of LMOs.

scholarly journals Development of a Multiplex PCR Assay to Monitor Living Modified Cottons in South Korea

2019 ◽  
Vol 9 (13) ◽  
pp. 2688
Author(s):  
Kim ◽  
Kim ◽  
Lim ◽  
Choi ◽  
Lee
Keyword(s):  
South Korea ◽  
Multiplex Pcr ◽  
Detection Method ◽  
Simultaneous Detection ◽  
Livestock Feed ◽  
Multiplex Pcr Assay ◽  
Distribution Process ◽  
Pcr Method ◽  
Living Modified Organisms ◽  
Quality Of Milk

Cotton has been cultivated worldwide and is a useful crop for humans. However, all living modified organisms (LMOs), including living modified (LM) cotton, are not cultivated in South Korea and are imported from overseas. LM cotton imports are on the rise and most of the imported cotton is used as livestock feed. In particular, it is commonly used to feed Holstein breeds that produce milk, because cotton improves the quality of milk. However, as the cotton imports increase, the possibility of unintentional outflows in the distribution process also increases. Consequently, there is an increasing concern about unintentional release of LM cotton into the natural environment. Therefore, environmental monitoring and post-management of LMOs are very important steps. Recently, a total of 30 LM crop events were approved for LM cotton import in South Korea. A single detection method has been used to monitor individual events. However, a single method of detection for collected samples requires a large number of PCRs, with obvious disadvantages. Therefore, a simultaneous detection method was developed for 8 representative events (GHB119, GHB614, MON88913, MON15985, LLCOTTON25, MON1445, 281-3006, and MON531) in an effort to monitor 26 of them and facilitate the identification of LM cotton. The results suggest that our new multiplex PCR method may be useful for monitoring and post-management of LM cotton.

A PCR method targeting internal transcribed spacers: the simultaneous detection of Babesia bigemina and Babesia bovis in cattle

2014 ◽  
Vol 59 (1) ◽  
Author(s):  
Junlong Liu ◽  
Guiquan Guan ◽  
Aihong Liu ◽  
Youquan Li ◽  
Hong Yin ◽  
...  
Keyword(s):  
Light Microscopy ◽  
Multiplex Pcr ◽  
Simultaneous Detection ◽  
Internal Transcribed Spacers ◽  
Babesia Bovis ◽  
Babesia Bigemina ◽  
Pcr Assay ◽  
Blood Samples ◽  
Multiplex Pcr Assay ◽  
Pcr Method

AbstractIn this study, two pairs of oligonucleotide primers were designed according to the nucleotide sequence of the internal transcribed spacers (ITSs) of Babesia bigemina and B. bovis isolates from China. The primers were used in a multiplex PCR to detect parasite DNA in blood samples from cattle. There was no cross reactions with B. ovata, B. major, B. sp. Kashi, Theileria annulata, T. sergenti, T. sinensis or normal bovine DNA. The sensitivity of multiplex PCR assay was 1 pg and 10 pg DNA for B. bigemina and B. bovis, respectively. A total of 260 field blood samples collected from cattle in five provinces of China were analyzed by multiplex PCR and light microscopy. PCR testing revealed that 7.3% (19/260) and 5.8% (15/260) of cattle were positive for B. bigemina and B. bovis and 1.2% (3/260) of cattle were co-infected with B. bigemina and B. bovis. Using light microscopy, 2.3% (6/260) and 1.5% (4/260) of cattle were infected by B. bigemina and B. bovis, respectively, and no co-infection was found. The results showed that the multiplex PCR developed in the present study could be an alternative diagnostic tool for the detection of B. bovis and B. bigemina infection in cattle.

Multiplex PCR assay to discriminate four neighbouring species of the Calliptamus genus (Orthoptera: Acrididae) from France

2010 ◽  
Vol 100 (6) ◽  
pp. 701-706 ◽  
Author(s):  
E. Blanchet ◽  
L. Blondin ◽  
P.A. Gagnaire ◽  
A. Foucart ◽  
J.M. Vassal ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Species Identification ◽  
Mitochondrial Cytochrome ◽  
Pcr Assay ◽  
Species Determination ◽  
Southern France ◽  
Multiplex Pcr Assay ◽  
Pcr Method ◽  
Definition Of ◽  
Cytochrome Oxydase

AbstractDefinition of the genus Calliptamus (Orthoptera: Acrididae) has generated many taxonomic debates. Even now, the existence of different geographical morphs hinders species determination, particularly as concerns females and larvae. Some of these species are observed in southern France and are recognized as potential pests. To circumvent problems of species identification in ecological surveys, we developed a single multiplex PCR method based on mitochondrial Cytochrome Oxydase I diagnostic polymorphisms to differentiate between the four species, Calliptamus italicus, C. wattenwylianus, C. siciliae and C. barbarus, in southern regions of France.

Identification of cytoplasm types in rapeseed (Brassica napus L.) accessions by a multiplex PCR assay

2010 ◽  
Vol 121 (4) ◽  
pp. 643-650 ◽  
Author(s):  
H. X. Zhao ◽  
Z. J. Li ◽  
S. W. Hu ◽  
G. L. Sun ◽  
J. J. Chang ◽  
...  
Keyword(s):  
Brassica Napus ◽  
Multiplex Pcr ◽  
Pcr Assay ◽  
Brassica Napus L ◽  
Multiplex Pcr Assay

Diagnostic Yield of Multiplex PCR Method in Cerebrospinal Fluid for the Diagnosis of Purulent Meningitis in Children

2020 ◽  
Author(s):  
Jing-Li Zhao ◽  
Chun-Zhen Hua ◽  
Yong-Ping Xie ◽  
Yan-Xiang Pan ◽  
Bo-Fei Hu ◽  
...  
Keyword(s):  
Cerebrospinal Fluid ◽  
Streptococcus Pneumoniae ◽  
Multiplex Pcr ◽  
Diagnostic Yield ◽  
Pcr Assay ◽  
E Coli ◽  
Purulent Meningitis ◽  
Multiplex Pcr Assay ◽  
Pcr Method ◽  
Bacteria Culture

Abstract Objective To evaluate the diagnostic yield of the multiplex polymerase chain reaction (PCR) method in cerebrospinal fluid (CSF) for the diagnosis of purulent meningitis (PM) in children. Methods PM was diagnosed according to the European Society for Clinical Microbiology and Infectious Diseases guideline (2016). Patients with PM between May 2015 and October 2018 were included. The multiplex PCR method was used to detect eight common identified bacteria in PM. Its sensitivity and specificity were compared with bacteria culture. Results A total of 106 cases were enrolled. Pathogenic bacteria were identified in 27 (25.5%) cases by culture and in 37 (34.9%) cases by multiplex PCR assay. The top three bacteria were Streptococcus pneumoniae, Escherichia coli K1, and Streptococcus agalactiae. When using culture as the gold standard, the multiplex PCR assay showed a sensitivity of 100, 88.9, and 75.0% for S. agalactiae, S. pneumoniae, and E. coli K1, respectively, and a specificity of more than 91.3% for all three bacteria. For detectable bacteria, the positive rate of the multiplex PCR assay (36.6%, 37/101) was significantly higher than that of the bacteria culture (21.8%, 22/101). When combining the two methods, etiology was identified in 42.5% (45/106) of the patients. Conclusion Streptococcus pneumoniae, E. coli K1, and S. agalactiae were the predominant pathogens causing pediatric PM. As a rapid method with high sensitivity and specificity, the multiplex PCR assay in CSF could be used as an adjunctive approach with bacteria culture for the pathogen identification of PM.

scholarly journals Simple and Reliable Multiplex PCR Assay for Surveillance Isolates of Vancomycin-Resistant Enterococci

2000 ◽  
Vol 38 (8) ◽  
pp. 3092-3095 ◽  
Author(s):  
Reiko Kariyama ◽  
Ritsuko Mitsuhata ◽  
Joseph W. Chow ◽  
Don B. Clewell ◽  
Hiromi Kumon
Keyword(s):  
Multiplex Pcr ◽  
Pcr Assay ◽  
Vancomycin Resistant Enterococci ◽  
Multiplex Pcr Assay ◽  
Pcr Method ◽  
Vancomycin Resistant ◽  
Primer Sets ◽  
Rapid Characterization ◽  
Surveillance Specimens

Conditions have been optimized for the use of a multiplex PCR for the detection of vancomycin-resistant enterococci in nosocomial surveillance specimens. Seven primer sets targeting the genesvanA, vanB, vanC1, vanC2/C3 Enterococcus faecalis-specific, Enterococcus faecium-specific, and rrs (16S rRNA) were used in one reaction tube. The PCR method developed in the present study is simple and reliable for the rapid characterization of vancomycin-resistant enterococci.

Usefulness of a Multiplex PCR for Detection of Diarrheagenic Escherichia coli in a Diagnostic Microbiology Laboratory Setting

2016 ◽  
Vol 1 (2) ◽  
pp. 38-42 ◽  
Author(s):  
Khairun Nessa ◽  
Dilruba Ahmed ◽  
Johirul Islam ◽  
FM Lutful Kabir ◽  
M Anowar Hossain
Keyword(s):  
Escherichia Coli ◽  
Multiplex Pcr ◽  
Clinical Laboratory ◽  
Proteinase K ◽  
Microbiology Laboratory ◽  
Pcr Assay ◽  
E Coli ◽  
Diarrheagenic Escherichia Coli ◽  
Multiplex Pcr Assay ◽  
Diagnostic Microbiology

A multiplex PCR assay was evaluated for diagnosis of diarrheagenic Escherichia coli in stool samples of patients with diarrhoea submitted to a diagnostic microbiology laboratory. Two procedures of DNA template preparationproteinase K buffer method and the boiling method were evaluated to examine isolates of E. coli from 150 selected diarrhoeal cases. By proteinase K buffer method, 119 strains (79.3%) of E. coli were characterized to various categories by their genes that included 55.5% enteroaggregative E. coli (EAEC), 18.5% enterotoxigenic E. coli (ETEC), 1.7% enteropathogenic E. coli (EPEC), and 0.8% Shiga toxin-producing E. coli (STEC). Although boiling method was less time consuming (<24 hrs) and less costly (<8.0 US $/ per test) but was less efficient in typing E. coli compared to proteinase K method (41.3% vs. 79.3% ; p<0.001). The sensitivity and specificity of boiling method compared to proteinase K method was 48.7% and 87.1% while the positive and negative predictive value was 93.5% and 30.7%, respectively. The majority of pathogenic E. coli were detected in children (78.0%) under five years age with 53.3% under one year, and 68.7% of the children were male. Children under 5 years age were frequently infected with EAEC (71.6%) compared to ETEC (24.3%), EPEC (2.7%) and STEC (1.4%). The multiplex PCR assay could be effectively used as a rapid diagnostic tool for characterization of diarrheagenic E. coli using a single reaction tube in the clinical laboratory setting.Bangladesh J Med Microbiol 2007; 01 (02): 38-42

New approach for the identification of potentially toxigenic Corynebacterium sp. using a multiplex PCR assay

2021 ◽  
pp. 106198
Author(s):  
Sunarno ◽  
Khariri ◽  
Fauzul Muna ◽  
Kambang Sariadji ◽  
Yuni Rukminiati ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Pcr Assay ◽  
New Approach ◽  
Multiplex Pcr Assay ◽  
Corynebacterium Sp

A multiplex PCR assay to diagnose and quantify Nosema infections in honey bees (Apis mellifera)

2010 ◽  
Vol 105 (2) ◽  
pp. 151-155 ◽  
Author(s):  
Mollah Md. Hamiduzzaman ◽  
Ernesto Guzman-Novoa ◽  
Paul H. Goodwin
Keyword(s):  
Apis Mellifera ◽  
Multiplex Pcr ◽  
Honey Bees ◽  
Pcr Assay ◽  
Multiplex Pcr Assay
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