Description of Perkinsus andrewsi n. sp. Isolated from the Baltic Clam (Macoma balthica) by Characterization of the Ribosomal RNA Locus, and Development of a Species-Specific PCR-Based Diagnostic Assay

2001 ◽  
Vol 48 (1) ◽  
pp. 52-61 ◽  
Author(s):  
CATHLEEN A. COSS ◽  
JOSE A. F. ROBLEDO ◽  
GREGORY M. RUIZ ◽  
GERARDO R. VASTA
Keyword(s):  
Ribosomal Rna ◽  
Macoma Balthica ◽  
Diagnostic Assay ◽  
Specific Pcr ◽  
The Baltic ◽  
Species Specific

scholarly journals Development and Validation of a Nested-PCR-Denaturing Gradient Gel Electrophoresis Method for Taxonomic Characterization of Bifidobacterial Communities

2003 ◽  
Vol 69 (11) ◽  
pp. 6380-6385 ◽  
Author(s):  
R. Temmerman ◽  
L. Masco ◽  
T. Vanhoutte ◽  
G. Huys ◽  
J. Swings
Keyword(s):  
Nested Pcr ◽  
Gel Electrophoresis ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Temporal Changes ◽  
Rrna Gene ◽  
Specific Pcr ◽  
Gradient Gel Electrophoresis ◽  
Species Specific ◽  
Taxonomic Characterization

ABSTRACT The taxonomic characterization of a bacterial community is difficult to combine with the monitoring of its temporal changes. None of the currently available identification techniques are able to visualize a “complete” community, whereas techniques designed for analyzing bacterial ecosystems generally display limited or labor-intensive identification potential. This paper describes the optimization and validation of a nested-PCR-denaturing gradient gel electrophoresis (DGGE) approach for the species-specific analysis of bifidobacterial communities from any ecosystem. The method comprises a Bifidobacterium-specific PCR step, followed by purification of the amplicons that serve as template DNA in a second PCR step that amplifies the V3 and V6-V8 regions of the 16S rRNA gene. A mix of both amplicons is analyzed on a DGGE gel, after which the band positions are compared with a previously constructed database of reference strains. The method was validated through the analysis of four artificial mixtures, mimicking the possible bifidobacterial microbiota of the human and chicken intestine, a rumen, and the environment, and of two fecal samples. Except for the species Bifidobacterium coryneforme and B. indicum, all currently known bifidobacteria originating from various ecosystems can be identified in a highly reproducible manner. Because no further cloning and sequencing of the DGGE bands is necessary, this nested-PCR-DGGE technique can be completed within a 24-h span, allowing the species-specific monitoring of temporal changes in the bifidobacterial community.

scholarly journals Isolation and Characterization of Brachyspira spp. from Dogs in the Czech Republic

2010 ◽  
Vol 79 (3) ◽  
pp. 437-442
Author(s):  
Daniel Šperling ◽  
František Čada ◽  
Alois Čížek
Keyword(s):  
Czech Republic ◽  
Biochemical Activity ◽  
Animal Shelters ◽  
The Czech Republic ◽  
Specific Pcr ◽  
Isolation And Characterization ◽  
High Prevalence ◽  
Species Specific

The objectives of this study were to establish the prevalence of intestinal Spirochetes of the genusBrachyspirain Czech dogs and to determine the susceptibility of obtainedB. pilosicoliisolates to selected antibacterial substances. Spirochetes were diagnosed microscopically in 23 out of 1139 samples of dogs’ excrements, primarily intended for a parasitological testing. The cultivation of positive samples provided 10 brachyspira isolates, which were, on the basis of their biochemical activity and the results of the species-specific PCR, identified asB. pilosicoli(9 isolates) andB. hyodysenteriae(1 isolate). These dogs came from households. All the 7 tested isolatesB. pilosicoliwere sensitive to metronidazole and doxycycline, uniformly resistant to erythromycin, partly sensitive to cefazoline, lincomicine and ampicilline except for one isolate ofB. pilosicoli, which was resistant to ampicilline. The second part of study was focused on dogs with diarrhoea that came from animal shelters, where a high prevalence of 58% (10/17) ofB. pilosicoliwas found.

scholarly journals Identification of a p28 Gene in Ehrlichia ewingii: Evaluation of Gene for Use as a Target for a Species-Specific PCR Diagnostic Assay

2001 ◽  
Vol 39 (11) ◽  
pp. 3871-3876 ◽  
Author(s):  
A. A. Gusa ◽  
R. S. Buller ◽  
G. A. Storch ◽  
M. M. Huycke ◽  
L. J. Machado ◽  
...  
Keyword(s):  
Diagnostic Assay ◽  
Specific Pcr ◽  
Species Specific

scholarly journals Molecular Characterization of Meloidogyne hispanica (Nematoda, Meloidogynidae) by Phylogenetic Analysis of Genes Within the rDNA in Meloidogyne spp.

Plant Disease ◽  
2008 ◽  
Vol 92 (7) ◽  
pp. 1104-1110 ◽  
Author(s):  
Blanca B. Landa ◽  
Juan E. Palomares Rius ◽  
Nicola Vovlas ◽  
Regina M. D. G. Carneiro ◽  
Carla M. N. Maleita ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Nematode Species ◽  
Maximum Parsimony Analysis ◽  
Biological Traits ◽  
Root Knot Nematode ◽  
Specific Pcr ◽  
Meloidogyne Spp ◽  
Species Specific ◽  
Prunus Spp

In the past, the distribution of Meloidogyne hispanica, the Seville root-knot nematode, appeared to be restricted to the southern part of Spain and Prunus spp.; however, its distribution has been confirmed to be worldwide because it occurs in all continents (Europe, Africa, Asia, Australia, and North, Central, and South America). Differentiation of M. hispanica from other Meloidogyne spp., mainly M. arenaria, can be very difficult using morphological and biological traits data. These species are quite similar and can be regularly confused in inaccurate taxonomic comparisons. In this study, species-specific polymerase chain reaction (PCR) and phylogenetic analysis of sequences from three ribosomal (r)DNA regions (18S, internal transcribed spacer [ITS]1-5.8S-ITS2, and D2-D3 of 28S) were used to characterize three M. hispanica isolates from different geographical origins (Brazil, Portugal, and Spain). Molecular analyses showed identical sequences for all three isolates for the three rDNA regions. Maximum parsimony analysis of the three rDNA regions and the species-specific PCR demonstrated and supported the differentiation of M. hispanica from M. incognita, M. javanica, and M. arenaria and from all described root-knot nematode species.

scholarly journals Molecular Characterization of the Toxic Cyanobacterium Cylindrospermopsis raciborskii and Design of a Species-Specific PCR

2000 ◽  
Vol 66 (1) ◽  
pp. 332-338 ◽  
Author(s):  
Kim M. Wilson ◽  
Mark A. Schembri ◽  
Peter D. Baker ◽  
Christopher P. Saint
Keyword(s):  
Cylindrospermopsis Raciborskii ◽  
Morphological Identification ◽  
Common Component ◽  
Climatic Regions ◽  
Specific Pcr ◽  
Species Specific ◽  
Genetic Profiles ◽  
Sequence Profiles ◽  
Toxic Bloom

ABSTRACT Cylindrospermopsis raciborskii is a toxic-bloom-forming cyanobacterium that is commonly found in tropical to subtropical climatic regions worldwide, but it is also recognized as a common component of cyanobacterial communities in temperate climates. Genetic profiles of C. raciborskii were examined in 19 cultured isolates originating from geographically diverse regions of Australia and represented by two distinct morphotypes. A 609-bp region of rpoC1, a DNA-dependent RNA polymerase gene, was amplified by PCR from these isolates with cyanobacterium-specific primers. Sequence analysis revealed that all isolates belonged to the same species, including morphotypes with straight or coiled trichomes. Additional rpoC1 gene sequences obtained for a range of cyanobacteria highlighted clustering of C. raciborskii with other heterocyst-producing cyanobacteria (orders Nostocales andStigonematales). In contrast, randomly amplified polymorphic DNA and short tandemly repeated repetitive sequence profiles revealed a greater level of genetic heterogeneity amongC. raciborskii isolates than did rpoC1 gene analysis, and unique band profiles were also found among each of the cyanobacterial genera examined. A PCR test targeting a region of therpoC1 gene unique to C. raciborskii was developed for the specific identification of C. raciborskiifrom both purified genomic DNA and environmental samples. The PCR was evaluated with a number of cyanobacterial isolates, but a PCR-positive result was only achieved with C. raciborskii. This method provides an accurate alternative to traditional morphological identification of C. raciborskii.

scholarly journals Detection And Molecular Characterization of Theileria Ovis in Sheep And Goats with Clinical The Ileriosis in Kurdistan, Iraq

2020 ◽  
Vol 23 (2) ◽  
pp. 69-78
Author(s):  
Zuber Ismael Hassen ◽  
Azad Abdullah Meerkhan
Keyword(s):  
Disease Transmission ◽  
Clinical Signs ◽  
Kurdistan Region ◽  
Single Test ◽  
Molecular Method ◽  
Blood Samples ◽  
Sheep And Goats ◽  
Specific Pcr ◽  
Species Specific

This study was carried out to detect Theileria infection in sheep and goats in Kurdistan region, Iraq from June 2019 to April 2020. Molecular method was used to identify Theileria species. Sixty- seven blood samples were taken from 45 sheep and 22 goats based on clinical signs of theileriosis during tick activating season. The 67 samples were PCR edm and as a result, 20 species-specific PCR were positives (26.67% (12/20) were Theileria ovis in sheep and 36.36% (8/20) were from goats). The results of the gene analysis in the current study were registered in NCBI under four accession numbers (MN889986, MN889987, MN889988 and MN889989), which shows that sheep and goats can be infected with Theileria ovis. This is the first report of Theileria species in goats with clinical theileriosis in Kurdistan, so the gene flow and disease transmission between sheep and goats is most expected. PCR is a useful diagnostic tool to detect ovine theileriosis with a single test and suggested that T. ovis is the dominant piroplasmid agent in Erbil. In addition, it revealed that sheep is very susceptible to theileriosis than goats

scholarly journals Isolation and Characterization of Mycoplasma mycoides Subspecies capri from Milk of Natural Goat Mastitis Cases

2013 ◽  
Vol 2013 ◽  
pp. 1-5 ◽  
Author(s):  
Vijay Kumar ◽  
Rajneesh Rana ◽  
Somya Mehra ◽  
Pramod Kumar Rout
Keyword(s):  
Restriction Analysis ◽  
Clinical Signs ◽  
Strain Variation ◽  
Specific Pcr ◽  
Isolation And Characterization ◽  
Mycoplasma Mycoides ◽  
Species Specific ◽  
Amplified Rdna Restriction Analysis ◽  
Indian Goat

Association of Mycoplasma mycoides subspecies capri (Mmc) with natural goat mastitis has been studied earlier largely by detecting the Mmc DNA using molecular methods. However, report on detection of cultivable Mmc isolates from natural goat-mastitis milk is still very rare. In this study, Mmc was isolated from milk samples () of goats with or without clinical signs of mastitis. Mmc isolates were further characterized by biochemical and species-specific PCR methods. Intra species strain variation was also studied by 16S amplified rDNA restriction analysis (16S ARDRA). The study recovered a total of 6 Mmc isolates (3.5%). Three types of intraspecies variants among the recovered Mmc isolates were found by 16S ARDRA. The study concluded that Mmc may be an etiological agent of mycoplasmal mastitis in Indian goat herds.

Molecular characterization of hospital-acquired adenovirus infantile respiratory infection in Chile using species-specific PCR assays

2007 ◽  
Vol 39 (3) ◽  
pp. 175-181 ◽  
Author(s):  
Carmen Larrañaga ◽  
Jorge Martínez H. ◽  
Angélica Palomino M. ◽  
Mónica Peña C. ◽  
Flavio Carrión A. ◽  
...  
Keyword(s):  
Molecular Characterization ◽  
Respiratory Infection ◽  
Specific Pcr ◽  
Pcr Assays ◽  
Species Specific ◽  
Hospital Acquired
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