A Variable 23S rDNA Region is a Useful Discriminating Target for Genus-Specific and Species-Specific PCR Amplification in Arcobacter Species

1995 ◽  
Vol 18 (3) ◽  
pp. 353-356 ◽  
Author(s):  
Katrin Bastyns ◽  
Dagmar Cartuyvels ◽  
Sabine Chapelle ◽  
Peter Vandamme ◽  
Herman Goossens ◽  
...  
Keyword(s):  
Pcr Amplification ◽  
Specific Pcr ◽  
Species Specific

A simple method for discriminating Bursaphelenchus xylophilus and B. mucronatus by species-specific polymerase chain reaction primer pairs

Nematology ◽  
2004 ◽  
Vol 6 (2) ◽  
pp. 273-277 ◽  
Author(s):  
Koji Matsunaga ◽  
Katsumi Togashi
Keyword(s):  
Pcr Amplification ◽  
Nematode Species ◽  
Bursaphelenchus Xylophilus ◽  
Polymerase Chain Reaction Primer ◽  
Dna Fragments ◽  
Simple Method ◽  
Specific Pcr ◽  
Pcr Products ◽  
Specific Pcr Primer ◽  
Species Specific

Abstract Two species-specific PCR primer pairs were developed for identifying the two nematode species, Bursaphelenchus xylophilus and B. mucronatus. The primer pairs were developed from the sequence of ribosomal DNA (rDNA) repeats to produce DNA fragments of different lengths by PCR amplification. The DNA fragments for B. mucronatus and B. xylophilus were 210 bp and 557 bp, respectively. When mixed, neither primer pair inhibited the PCR amplification of the other. Five isolates of B. xylophilus and four isolates of B. mucronatus showed different band profiles of PCR products between the two species, but identical profiles among isolates of the same species.

Specific PCR Detection of Arcobacter butzleri, Arcobacter cryaerophilus, Arcobacter skirrowii, and Arcobacter cibarius in Chicken Meat

2009 ◽  
Vol 72 (7) ◽  
pp. 1491-1495 ◽  
Author(s):  
DANIELA PENTIMALLI ◽  
NICOLETTE PEGELS ◽  
TERESA GARCÍA ◽  
ROSARIO MARTÍN ◽  
ISABEL GONZÁLEZ
Keyword(s):  
Pcr Amplification ◽  
Chicken Meat ◽  
Pcr Analysis ◽  
Rrna Gene ◽  
Pcr Assay ◽  
Gyra Gene ◽  
Specific Primers ◽  
Specific Pcr ◽  
Arcobacter Butzleri ◽  
Species Specific

An enrichment PCR assay using species-specific primers was developed for the detection of Arcobacter butzleri, Arcobacter cryaerophilus, Arcobacter skirrowii, and Arcobacter cibarius in chicken meat. Primers for A. cryaerophilus, A. skirrowii, and A. cibarius were designed based on the gyrA gene to amplify nucleic acid fragments of 212, 257, and 145 bp, respectively. The A. butzleri–specific primers were designed flanking a 203-bp DNA fragment in the 16S rRNA gene. The specificity of the four primer pairs was assessed by PCR analysis of DNA from a panel of Arcobacter species, related Campylobacter, Helicobacter species, and other food bacteria. The applicability of the method was then validated by testing 42 fresh retail-purchased chicken samples in the PCR assay. An 18-h selective preenrichment step followed by PCR amplification with the four Arcobacter primer sets revealed the presence of Arcobacter spp. in 85.7% of the retail chicken samples analyzed. A. butzleri was the only species present in 50% of the samples, and 35.7% of the samples were positive for both A. butzleri and A. cryaerophilus. A. skirrowii and A. cibarius were not detected in any of the chicken samples analyzed. The enrichment PCR assay developed is a specific and rapid alternative for the survey of Arcobacter contamination in meat.

scholarly journals Species-Specific PCR Assays for Differentiating Heterodera filipjevi and H. avenae

Plant Disease ◽  
2013 ◽  
Vol 97 (12) ◽  
pp. 1611-1619 ◽  
Author(s):  
Guiping Yan ◽  
Richard W. Smiley ◽  
Patricia A. Okubara ◽  
Andrea M. Skantar
Keyword(s):  
Pacific Northwest ◽  
Management Practices ◽  
Pcr Amplification ◽  
Nematode Species ◽  
Heterodera Avenae ◽  
Cyst Nematodes ◽  
Specific Pcr ◽  
The Pacific ◽  
Pcr Assays ◽  
Species Specific

Heterodera avenae and H. filipjevi are economically important cyst nematodes that restrict production of cereal crops in the Pacific Northwest United States and elsewhere in the world. Identification of these two species is critical for recommending and implementing effective management practices. Primers were designed from the internal transcribed spacer (ITS) regions of H. avenae and H. filipjevi ribosomal DNA. The primers were highly specific when examined on target isolates but did not amplify DNA from nontarget Heterodera, Globodera, Meloidogyne, Pratylenchus, and other nematode species tested. Polymerase chain reaction (PCR) and amplification conditions were established, and H. avenae and H. filipjevi were clearly distinguished by PCR fragments of 242 and 170 bp, respectively. Robust PCR amplification was achieved with DNA extracted from a single egg or second-stage juvenile (J2) using a laboratory-made worm lysis buffer, and DNA from 0.5 egg or J2 using a commercial kit. The PCR assays were successfully employed for differentiation of H. filipjevi and H. avenae populations collected from eight locations in three Pacific Northwest states. This is the first report of a species-specific ITS PCR assay to detect and identify H. filipjevi. The assays for both species will enhance diagnosis of cereal cyst nematode species in infested fields.

scholarly journals Species-Specific Plasmid Sequences for PCR Identification of the Three Species of Borrelia burgdorferiSensu Lato Involved in Lyme Disease

1998 ◽  
Vol 36 (1) ◽  
pp. 269-272 ◽  
Author(s):  
Marie-Christine Misonne ◽  
Philippe Pierre Hoet
Keyword(s):  
Lyme Disease ◽  
Pcr Amplification ◽  
Sensu Stricto ◽  
Pcr Assay ◽  
Borrelia Garinii ◽  
Specific Pcr ◽  
Pcr Identification ◽  
Identification Of Species ◽  
Species Specific ◽  
Specific Sequences

Species-specific sequences were shown to be carried by plasmids of the three main species of Borrelia burgdorferi sensu lato involved in Lyme disease. Libraries of the 16-, 33-, and 25-kb plasmids of B. burgdorferi sensu stricto, Borrelia garinii, and Borrelia afzelii, respectively, were then built and used to isolate species-specific sequences. After sequencing of the cloned inserts, three sets of primers were designed. They were shown to determine species-specific PCR amplification products. The sensitivities of the PCR assay with these primers were 100 spirochetes for B. burgdorferi sensu stricto and 1,000 spirochetes for B. garinii and B. afzelii. The usefulness of these primers for the identification of species in biological samples (tick, serum, and cerebrospinal fluid samples) was ascertained.

Pigment-cell-specific genes from fibroblasts are transactivated after chromosomal transfer into melanoma cells

1994 ◽  
Vol 14 (2) ◽  
pp. 1179-1190
Author(s):  
T P Powers ◽  
T B Shows ◽  
R L Davidson
Keyword(s):  
Pigment Cell ◽  
Pcr Amplification ◽  
Melanoma Cells ◽  
Mouse Fibroblast ◽  
B Locus ◽  
Specific Pcr ◽  
Tyrosinase Gene ◽  
Microcell Hybrid ◽  
Microcell Hybrids ◽  
Species Specific

Human and mouse fibroblast chromosomes carrying tyrosinase or b-locus genes were introduced, by microcell hybridization, into pigmented Syrian hamster melanoma cells, and the microcell hybrids were tested for transactivation of the fibroblast tyrosinase and b-locus genes. By using species-specific PCR amplification to distinguish fibroblast and melanoma cDNAs, it was demonstrated that the previously silent fibroblast tyrosinase and b-locus genes were transactivated following chromosomal transfer into pigmented melanoma cells. However, transactivation of the mouse fibroblast tyrosinase gene was unstable in microcell hybrid subclones and possibly dependent on a second fibroblast locus that could have segregated in the subclones. This second locus was not necessary for transactivation of the fibroblast b-locus gene, thus demonstrating noncoordinate transactivation of fibroblast tyrosinase and b-locus genes. Transactivation of the fibroblast tyrosinase gene in microcell hybrids apparently is dependent on the absence of a putative fibroblast extinguisher locus for tyrosinase gene expression, which presumably is responsible for the extinction of pigmentation in hybrids between karyotypically complete fibroblasts and melanoma cells.

scholarly journals Pigment-cell-specific genes from fibroblasts are transactivated after chromosomal transfer into melanoma cells.

1994 ◽  
Vol 14 (2) ◽  
pp. 1179-1190 ◽  
Author(s):  
T P Powers ◽  
T B Shows ◽  
R L Davidson
Keyword(s):  
Pigment Cell ◽  
Pcr Amplification ◽  
Melanoma Cells ◽  
Mouse Fibroblast ◽  
B Locus ◽  
Specific Pcr ◽  
Tyrosinase Gene ◽  
Microcell Hybrid ◽  
Microcell Hybrids ◽  
Species Specific

Human and mouse fibroblast chromosomes carrying tyrosinase or b-locus genes were introduced, by microcell hybridization, into pigmented Syrian hamster melanoma cells, and the microcell hybrids were tested for transactivation of the fibroblast tyrosinase and b-locus genes. By using species-specific PCR amplification to distinguish fibroblast and melanoma cDNAs, it was demonstrated that the previously silent fibroblast tyrosinase and b-locus genes were transactivated following chromosomal transfer into pigmented melanoma cells. However, transactivation of the mouse fibroblast tyrosinase gene was unstable in microcell hybrid subclones and possibly dependent on a second fibroblast locus that could have segregated in the subclones. This second locus was not necessary for transactivation of the fibroblast b-locus gene, thus demonstrating noncoordinate transactivation of fibroblast tyrosinase and b-locus genes. Transactivation of the fibroblast tyrosinase gene in microcell hybrids apparently is dependent on the absence of a putative fibroblast extinguisher locus for tyrosinase gene expression, which presumably is responsible for the extinction of pigmentation in hybrids between karyotypically complete fibroblasts and melanoma cells.

scholarly journals Species-specific RT-PCR amplification of human enteroviruses: a tool for rapid species identification of uncharacterized enteroviruses

2006 ◽  
Vol 87 (1) ◽  
pp. 119-128 ◽  
Author(s):  
M. Steven Oberste ◽  
Kaija Maher ◽  
Alford J. Williams ◽  
Naomi Dybdahl-Sissoko ◽  
Betty A. Brown ◽  
...  
Keyword(s):  
Pcr Amplification ◽  
Pcr Primers ◽  
Human Enterovirus ◽  
Rt Pcr ◽  
Specific Primer ◽  
Specific Primers ◽  
Typical Application ◽  
Partial Sequencing ◽  
Specific Pcr ◽  
Species Specific

The 65 serotypes of human enteroviruses are classified into four species, Human enterovirus (HEV) A to D, based largely on phylogenetic relationships in multiple genome regions. The 3′-non-translated region of enteroviruses is highly conserved within a species but highly divergent between species. From this information, species-specific RT-PCR primers were developed that can be used to rapidly screen collections of enterovirus isolates to identify species of interest. The four primer pairs were 100 % specific when tested against enterovirus prototype strains and panels of isolates of known serotype (a total of 193 isolates). For evaluation in a typical application, the species-specific primers were used to screen 186 previously uncharacterized non-polio enterovirus isolates. The HEV-B primers amplified 68·3 % of isolates, while the HEV-A and HEV-C primers accounted for 9·7 and 11·3 % of isolates, respectively; no isolates were amplified with the HEV-D primers. Twelve isolates (6·5 %) were amplified by more than one primer set and eight isolates (4·3 %) were not amplified by any of the four primer pairs. Serotypes were identified by partial sequencing of the VP1 capsid gene, and in every case sequencing confirmed that the species-specific PCR result was correct; the isolates that were amplified by more than one species-specific primer pair were mixtures of two (11 isolates) or three (one isolate) species of viruses. The eight isolates that were not amplified by the species-specific primers comprised four new serotypes (EV76, EV89, EV90 and EV91) that appear to be unique members of HEV-A based on VP1, 3D and 3′-non-translated region sequences.

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