Use of an arbitrarily primed PCR product in the development of a Campylobacter jejuni-specific PCR.

1997 ◽  
Vol 63 (3) ◽  
pp. 1019-1023 ◽  
Author(s):  
W A Day ◽  
I L Pepper ◽  
L A Joens
Keyword(s):  
Campylobacter Jejuni ◽  
Specific Pcr ◽  
Pcr Product ◽  
Arbitrarily Primed Pcr

scholarly journals The polymorphism of ETR1 gene in autochthonous apple cultivars

Genetika ◽  
2007 ◽  
Vol 39 (3) ◽  
pp. 387-394 ◽  
Author(s):  
Sladjana Maric ◽  
Radovan Boskovic ◽  
Milan Lukic
Keyword(s):  
Plant Hormone ◽  
Allelic Polymorphism ◽  
Ethylene Receptor ◽  
Gene Encoding ◽  
Fruit Storage ◽  
Specific Pcr ◽  
Pcr Product ◽  
Apple Cultivars ◽  
Storage Ability

Ethylene is a plant hormone, which plays an important role in the ripening of climacteric fruits such as the apple. We studied allelic polymorphism of the ETR1 gene, encoding ethylene receptor, in 23 autochthonous apple cultivars. The polymorphism was revealed by combining the gene specific PCR and restriction of PCR product. Four alleles of the ETR1 gene (a, b, c and d) were detected, and their possible association with the fruit storage ability examined.

scholarly journals Rapid Detection Method of Salmonella-specific PCR Product by DNA-immobilized Quartz Crystal Microbalance.

1999 ◽  
Vol 16 (1) ◽  
pp. 57-63
Author(s):  
Takahisa MIYAMOTO ◽  
Sudsai TREVANICH ◽  
Takashi OKABE ◽  
Satosi TOMODA ◽  
Ken-ichi HONJOH ◽  
...  
Keyword(s):  
Quartz Crystal Microbalance ◽  
Rapid Detection ◽  
Quartz Crystal ◽  
Detection Method ◽  
Specific Pcr ◽  
Pcr Product

Prognosis of glioblastoma with faint MGMT methylation-specific PCR product

2015 ◽  
Vol 122 (1) ◽  
pp. 179-188 ◽  
Author(s):  
Chih-Yi Hsu ◽  
Hsiang-Ling Ho ◽  
Shih-Chieh Lin ◽  
Yi-Chun Chang-Chien ◽  
Ming-Hsiung Chen ◽  
...  
Keyword(s):  
Mgmt Methylation ◽  
Methylation Specific Pcr ◽  
Specific Pcr ◽  
Pcr Product

scholarly journals Multiplex-Polymerase Chain Reaction Assay for the Authentication of the Mackerel Scomber colias in Commercial Canned Products

2006 ◽  
Vol 89 (3) ◽  
pp. 708-711 ◽  
Author(s):  
Carlos Infante ◽  
Manuel Manchado
Keyword(s):  
Polymerase Chain Reaction ◽  
Multiplex Polymerase Chain Reaction ◽  
Rrna Gene ◽  
Chain Reaction ◽  
Nontranscribed Spacer ◽  
Specific Pcr ◽  
Scomber Colias ◽  
Pcr Product ◽  
Polymerase Chain ◽  
Specific Fragment

Abstract A multiplex-polymerase chain reaction (PCR) system was developed for the authentication of the mackerel Scomber colias in commercial canned products. This novel method consists of an S. colias-specific fragment [159 base pairs (bp)] located in the nontranscribed spacer (NTS) sequence, and a Scomber genus-specific PCR product in the 5S rRNA gene (196201 bp) as a positive amplification control. The system was assayed using 18 different canned products labeled as S. colias. A positive identification was made in all but one sample, revealing this methodology as a potential molecular tool for direct application in the authentication of S. colias canned products.

scholarly journals Prevalence of Pandemic Thermostable Direct Hemolysin-Producing Vibrio parahaemolyticus O3:K6 in Seafood and the Coastal Environment in Japan

2003 ◽  
Vol 69 (7) ◽  
pp. 3883-3891 ◽  
Author(s):  
Yukiko Hara-Kudo ◽  
Kanji Sugiyama ◽  
Mitsuaki Nishibuchi ◽  
Ashrafuzzaman Chowdhury ◽  
Jun Yatsuyanagi ◽  
...  
Keyword(s):  
Vibrio Parahaemolyticus ◽  
The United States ◽  
Most Probable Number ◽  
Asian Countries ◽  
Probable Number ◽  
Specific Pcr ◽  
Thermostable Direct Hemolysin ◽  
Chromogenic Agar ◽  
Pcr Method ◽  
Arbitrarily Primed Pcr

ABSTRACT Although thermostable direct hemolysin (TDH)-producing Vibrio parahaemolyticus has caused many infections in Asian countries, the United States, and other countries, it has been difficult to detect the same pathogen in seafoods and other environmental samples. In this study, we detected and enumerated tdh gene-positive V. parahaemolyticus in Japanese seafoods with a tdh-specific PCR method, a chromogenic agar medium, and a most-probable-number method. The tdh gene was detected in 33 of 329 seafood samples (10.0%). The number of tdh-positive V. parahaemolyticus ranged from <3 to 93/10 g. The incidence of tdh-positive V. parahaemolyticus tended to be high in samples contaminated with relatively high levels of total V. parahaemolyticus. TDH-producing strains of V. parahaemolyticus were isolated from 11 of 33 tdh-positive samples (short-necked clam, hen clam, and rock oyster). TDH-producing strains of V. parahaemolyticus were also isolated from the sediments of rivers near the coast in Japan. Representative strains of the seafood and sediment isolates were examined for the O:K serovar and by the PCR method specific to the pandemic clone and arbitrarily primed PCR and pulsed-field gel electrophoresis techniques. The results indicated that most O3:K6 tdh-positive strains belonged to the pandemic O3:K6 clone and suggested that serovariation took place in the Japanese environment.

scholarly journals Allele-Specific PCR for the Detection of Azoxystrobin Resistance in Didymella bryoniae

Plant Disease ◽  
2014 ◽  
Vol 98 (12) ◽  
pp. 1681-1684 ◽  
Author(s):  
Mavis J. Finger ◽  
Venkatesan Parkunan ◽  
Pingsheng Ji ◽  
Katherine L. Stevenson
Keyword(s):  
Dna Sequences ◽  
Cyt B ◽  
Didymella Bryoniae ◽  
Specific Pcr ◽  
Pcr Product ◽  
Allele Specific ◽  
Polymerase Chain ◽  
Sensitive Allele ◽  
Cyt B Gene ◽  
Allele Specific Pcr

Gummy stem blight (GSB), caused by the fungus Didymella bryoniae, is considered the most widespread and destructive disease of watermelon in the southeastern United States. The quinone outside-inhibiting (QoI) fungicide azoxystrobin (AZO), which inhibits mitochondrial respiration by binding to the outer, quinone-oxidizing pocket of the cytochrome bc1 (cyt b) enzyme complex, was initially very effective in controlling GSB. However, resistance to AZO has been observed in D. bryoniae in many watermelon-producing regions. In this study, the DNA sequences of partial cyt b genes of four AZO-resistant (AZO-R) and four AZO-sensitive (AZO-S) isolates of D. bryoniae confirmed the amino acid substitution of glycine by alanine at the 143 codon (G143A) in the AZO-R isolates tested. Allele-specific primers were designed to detect the resistant or sensitive allele at codon 143 of the cyt b gene, which amplified a 165-bp polymerase chain reaction (PCR) product from genomic DNA of nine AZO-R and nine AZO-S isolates of D. bryoniae, respectively. The primer pairs did not amplify DNA from other pathogens tested in the study. The results indicated that the PCR assays developed in the study were specific in differentiating AZO-R and AZO-S isolates and could facilitate AZO resistance detection in D. bryoniae.

scholarly journals Genetic and Morphological Characterization of Colletotrichum acutatum Causing Anthracnose of Lupins

2002 ◽  
Vol 92 (9) ◽  
pp. 986-996 ◽  
Author(s):  
Pedro Talhinhas ◽  
S. Sreenivasaprasad ◽  
João Neves-Martins ◽  
Helena Oliveira
Keyword(s):  
Morphological Characters ◽  
Morphological Characterization ◽  
Colletotrichum Acutatum ◽  
Distinct Subgroup ◽  
Specific Pcr ◽  
Pathogen Diversity ◽  
Polymerase Chain ◽  
Arbitrarily Primed Pcr ◽  
Pathogen Diagnosis

Anthracnose, caused by Colletotrichum sp., is a serious problem of lupins (Lupinus spp.) worldwide. Morphological characters and molecular markers were used to characterize 43 Colletotrichum isolates from lupins, 8 isolates from other hosts, and 18 reference isolates representing related Colletotrichum spp., to assess the pathogen diversity and resolve its taxonomy. All lupin Colletotrichum isolates tested positive with C. acutatum-specific polymerase chain reaction (PCR) and did not test positive with C. gloeosporioides-specific PCR. Spore shape and colony diameter as well as insensitivity to benomyl grouped the lupin anthracnose isolates closer to C. acutatum than to C. gloeosporioides. Analysis of internal transcribed spacer (ITS) sequences of 57 Colletotrichum isolates grouped all lupin isolates with C. acutatum and distinct from C. gloeosporioides. Further, tub2 and his4 sequences revealed groups concordant with ITS, reducing the excessive dependence on the latter. Arbitrarily primed-PCR and amplified fragment length polymorphism analyses revealed intraspecific subgroups, but neither was useful to decipher species level relationships. ITS, tub2, and his4 results strongly support designating lupin anthracnose pathogen as C. acutatum or its subspecies. Most Colletotrichum isolates from lupins from worldwide locations are genetically homogeneous and form a distinct subgroup within C. acutatum. Present results also underline the potential of the C. acutatum-specific PCR for routine pathogen diagnosis.

PCR Identification of Ruminant Tissue in Raw and Heat-Treated Meat Meals

2006 ◽  
Vol 69 (9) ◽  
pp. 2241-2247 ◽  
Author(s):  
JEONG CHUL HA ◽  
WAN TAE JUNG ◽  
YONG SUK NAM ◽  
TAE WHA MOON
Keyword(s):  
Dna Sequence ◽  
12S Rrna ◽  
Spongiform Encephalopathy ◽  
Rrna Gene ◽  
Specific Pcr ◽  
Pcr Product ◽  
Heat Treated ◽  
Mitochondrial 12S Rrna ◽  
Animal Feedstuffs ◽  
Species Specific

To control the spread of bovine spongiform encephalopathy in cattle through contaminated animal feedstuffs, screening of feed products is essential. We designed five pairs of primers to identify specifically raw and heat-treated tissue from cattle, sheep, goat, deer, and ruminants in general. A forward common primer was designed based on a conserved DNA sequence in the mitochondrial 12S rRNA–tRNAval–16S rRNA gene, and reverse primers were designed to hybridize with a species-specific DNA sequence for each species considered. All primers were developed to create a specific PCR product small enough (less than 200 bp) to be suitable for heat-treated material. To evaluate the effect of heat treatment, a severe sterilization condition (133°C at 300 kPa for 20 min) was chosen. Species-specific amplicons were obtained from all types of heat-treated meat meals. Analysis of laboratory-contaminated vegetable meals revealed that the detection limit of the assay was 0.05% for each species analyzed. This PCR-based analysis can be used as a routine method for detecting banned animal-derived ingredients in raw and heat-treated feedstuffs.

scholarly journals Establishment of Methylation-Specific PCR for the Mouse p53 Gene

2011 ◽  
Vol 2011 ◽  
pp. 1-4 ◽  
Author(s):  
Ryuji Okazaki ◽  
Akira Ootsuyama ◽  
Yasuhiro Yoshida ◽  
Toshiyuki Norimura
Keyword(s):  
Cpg Islands ◽  
P53 Gene ◽  
Pcr Primers ◽  
Young Mouse ◽  
Methylation Specific Pcr ◽  
Specific Pcr ◽  
Pcr Product ◽  
Induced Tumors ◽  
Radiation Induced ◽  
Radiation Induced Tumors

Methylation-specific PCR (MSP) of the mouse p53 gene has not yet been reported. We searched the CpG islands, sequenced the bisulfited DNA, and designed PCR primers for methylation and unmethylation sites. DNA from a young mouse produced a strong PCR product with the unmethylated primer and a weaker band with the methylated primer. DNA from an old mouse produced bands of similar intensities with both primers. In radiation-induced tumors, DNA from an old mouse yielded similar bands with both types of primers. We suggest that MSP is a valuable technique for the epigenetic study of the mouse p53 gene.

scholarly journals Studying The Expression of Human P53 Gene in Patients Carrying Breast Cancer.

2017 ◽  
Vol 28 (1) ◽  
pp. 14
Author(s):  
Dawood S. Edan
Keyword(s):  
Breast Cancer ◽  
Healthy Woman ◽  
P53 Gene ◽  
Suppressor Gene ◽  
Women Patients ◽  
Cell Protection ◽  
Specific Pcr ◽  
Pcr Product ◽  
Dna Repair Proteins ◽  
Healthy Sample

The gene P53 can activate DNA repair proteins when DNA has sustained damage. So, it is an important factor in aging. Also, P53 gene can also be modified by mutagens (chemicals, radiation, or viruses), increasing the likelihood for uncontrolled cell division. For this importance, we collected thirty samples from Yarmouk hospital in Baghdad, Iraq. Twenty-nine from women patients carrying breast cancer and the last one was from healthy woman, and the samples that we have taken were embedded in paraffin wax. The extracted RNA from each samples was used to check the expression of a suppressor gene (P53) via Real time PCR. We designed primers of P53 gene on encoded sequence (Exon), to make sure generation a specific PCR product of P53 gene by converting mRNA to cDNA then PCR Product. The expression of each samples was fluctuated between 0.1 fold to 0.2 fold compared with the control (the healthy sample). The highest expression showed was samples 7 (0.2 fold), and the lowest expression showed was samples 27 (0.1 fold) that compared with control expression (sample 19). While the control expression was the highest (0.25 fold) among all samples. The results indicate that tumor may affects on a suppressor gene (P53) and can decrease the gene expression, and eventually can decrease the P53 suppressor protein that can be used it in the cell protection against cancer.

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