Dataset Information

Taxonomic Affinities and Intra- and Interspecific Variation in Bursaphelenchus spp. as Determined by Polymerase Chain Reaction.

ABSTRACT: Identification of closely related nematode species or races can be very difficult when diagnostic characters are plastic and overlapping. In this study we describe the use of polymerase chain reaction technology and direct DNA sequencing on 19 populations of Bursaphelenchus spp. to help understand their taxonomic relationships. The 5' end of the heat shock 70A gene from Caenorhabditis elegans was used as the target DNA sequence because it contains both coding and non-coding regions. The results indicate that the 19 populations could be divided into five types within B. xylophilus and four types within B. mucronatus. On a larger scale, the data revealed three distinct groups, representing B. xylophilus from North America and Japan, B. mucronatus from Japan, and "B. mucronatus" from Europe. There is sufficient difference between the European and Japanese "B. mucronatus" groups to warrant their consideration as separate species.

SUBMITTER: Beckenbach K

PROVIDER: S-EPMC2619233 | biostudies-other | 1992 Mar

REPOSITORIES: biostudies-other

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Taxonomic Affinities and Intra- and Interspecific Variation in Bursaphelenchus spp. as Determined by Polymerase Chain Reaction.

Beckenbach K K Smith M J MJ Webster J M JM

Journal of nematology 19920301 1

Identification of closely related nematode species or races can be very difficult when diagnostic characters are plastic and overlapping. In this study we describe the use of polymerase chain reaction technology and direct DNA sequencing on 19 populations of Bursaphelenchus spp. to help understand their taxonomic relationships. The 5' end of the heat shock 70A gene from Caenorhabditis elegans was used as the target DNA sequence because it contains both coding and non-coding regions. The results ...[more]

PMID: 19283215

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Project description:Background and aimDifferent polymerase chain reaction (PCR) techniques have and are still being used for the direct detection of Brucella DNA in serum samples of different animal species and humans without being validated or properly validated, resulting in discrepancies. Thus, this study aimed to evaluate the diagnostic performance of the TaqMan Real-Time-PCR (RT-PCR) targeting the bcsp31 gene versus conventional PCR for the accurate diagnosis of brucellosis at the genus level in cattle sera.Materials and methodsOne hundred and eighty-four serum samples were collected from bacteriologically positive and negative cows with ages ranging from 1 to 5 years old at some infected private farms in the Nile Delta under quarantine measures as well as brucellosis free farms. These samples were classified into four groups after serological diagnosis and investigated by TaqMan RT-PCR and conventional PCR targeting the IS711 gene for Brucella DNA detection. The diagnostic performance characteristics of both PCR techniques were estimated considering the bacteriological results as a gold standard.ResultsTaqMan RT-PCR revealed superiority over conventional PCR; it was able to detect Brucella DNA in 95% (67/70) and 89% (25/28) of the cattle sera samples belonging to Group 1 (serologically and bacteriologically positive) and Group 2 (serologically negative but bacteriologically positive), respectively. On evaluating the diagnostic performance, TaqMan RT-PCR showed superior diagnostic sensitivity (93.9%), diagnostic specificity (88.4%), performance index (182.3), almost perfect kappa agreement (0.825±0.042), strong positive correlation (r=0.826), high accuracy based on the receiver operating characteristic (ROC) curve, and area under the ROC curve (0.911) at p<0.05 and CI of 95%.ConclusionA cattle serum sample is not the metric of choice for targeting Brucella genomic DNA by conventional PCR. The time-saving and rapid TaqMan RT-PCR method revealed a better diagnostic performance in the detection of Brucella DNA in cattle sera. Such performance offered by TaqMan RT-PCR may be considered a step toward the possibility of using such technology in the direct differentiation between Brucella-infected and -vaccinated cattle immunized by smooth vaccines from cattle sera using primers specific for such vaccines.

Dataset Information

Taxonomic Affinities and Intra- and Interspecific Variation in Bursaphelenchus spp. as Determined by Polymerase Chain Reaction.

Publications

Taxonomic Affinities and Intra- and Interspecific Variation in Bursaphelenchus spp. as Determined by Polymerase Chain Reaction.

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