scholarly journals The application of genotyping techniques to the epidemiological analysis ofCampylobacter jejuni

1996 ◽  
Vol 117 (2) ◽  
pp. 233-244 ◽  
Author(s):  
C. J. Jackson ◽  
A. J. Fox ◽  
D. R. A. Wareing ◽  
D. N. Hutchinson ◽  
D. M. Jones

SummaryCampylobacter jejuniserogroup reference strains and collections of sporadic and outbreak- associated isolates were examined for restriction fragment length polymorphisms (RFLPs), usingC. jejunirandom chromosomal and 16S rRNA gene probes. A collection of 48 Penner (HS) and 14 Lior (HL) serogroup reference strains, plus 10 clinical isolates, generated 35 RFLP and 26 ribotype patterns. In combination the two loci generated 48 distinct genotypes. Both probes were able to differentiate between certain random isolates of the same HS/HL serogroups but greater discrimination was obtained with RFLP than with ribotyping. Genotyping distinguished accurately between related and unrelated strains when applied to several outbreaks. Genotypic analysis ofC. jejuniby restriction fragment length polymorphisms is a valuable technique for epidemiological typing. Chromosomal variation detected by the two unlinked probe loci provides some information about the genetic relationship between isolates.

1989 ◽  
Vol 35 (12) ◽  
pp. 1146-1155 ◽  
Author(s):  
J. Stephen Horton ◽  
Paul A. Horgen

Within the genus Achyla, which belongs to the class of fungi known as the Oomycetes, taxonomic judgments have traditionally been made using a variety of sexual criteria. We have used restriction fragment length polymorphisms as a new taxonomic character to examine intra- and inter-specific variation within this genus. Using a cDNA clone coding for the Achlya 18S rRNA gene as a hybridization probe, a 10-kb fragment of ribosomal DNA (rDNA) from Achlya ambisexualis strain E87 was cloned and then mapped for selected restriction enzyme sites. In Southern blot hybridizations, both this rDNA fragment and cloned 18S cDNAs revealed differences in the rDNA organization of A. ambisexualis E87 (male) and a female isolate of A. ambisexualis strain 734. No differences in the rDNAs were detected between the two heterothallic isolates A. ambisexualis E87 and A. bisexualis 65-1. Southern blot hybridizations suggested that two different rDNA organizations may exist within the genome of the homothallic strain A. heterosexualis B14. cDNA clones coding for two different hormonally regulated genes revealed the same relationships between the four isolates studied as those determined with rDNA probes. Two homothallic Achlya strains recently isolated from nature were found to have additional DNA polymorphisms not detected in the laboratory strains. Phenetic analysis distinguished the same similarities that were evident upon inspection of the hybridization data. Taken together, these data suggest different relationships between the isolates examined than do the previous taxonomic criteria by which species have been delimited within this genus.Key words: Achlya; restriction fragment length polymorphisms; ribosomal DNA; taxonomy.


1995 ◽  
Vol 114 (3) ◽  
pp. 423-431 ◽  
Author(s):  
A. P. Burnens ◽  
J. Wagner ◽  
H. Lior ◽  
J. Nicolet ◽  
J. Frey

SUMMARYSeveral typing systems have been described forCampylobacter jejuniandC. coli, to assess the complex epidemiology of these important enteric pathogens. In the present study two typing methods, slide agglutination according to the Lior scheme, and the demonstration of restriction-fragment length polymorphisms (RFLP) of flagellar genes, have been used in parallel on a set of 194 strains. This set comprised 118 sero-reference strains ofC. jejuniandC. coliof the Lior scheme, as well as 76 clinical isolates. All isolates were serotyped and subjected to PCR for amplification of flagellar genes, and the PCR product was restricted withAluI. Flagellar genes could be amplified in 152 strains. Among 85 seroreference strains, 74 different RFLP patterns were observed, and among 67 clinical isolates, there were 36 patterns. There was only limited correlation between flagellar RFLP and the Lior serogroup, and the variability of patterns in serogroups HL2 and HL4 were as marked as the variability between serogroups. Flagellar gene RFLP patterns are shown to be stable, highly discriminatory epidemiologic markers.


1988 ◽  
Vol 60 (02) ◽  
pp. 178-181 ◽  
Author(s):  
B R Bahnak ◽  
J M Lavergne ◽  
C L Verweij ◽  
C Rothschild ◽  
H Pannekoek ◽  
...  

SummaryDNA from a family with a female member affected with severe (type III) vWD was analysed using three restriction enzymes and a partial vWF cDNA probe. Two restriction fragment length polymorphisms (RFLPs) detected with the enzymes Bgl II and Xba I proved to be informative in this family. A 36.0 Kb allele, demonstrated with the enzyme Xba I was rare in the general population but very important in this family for segregation analysis of the alleles and their association with the putative defective chromosome. The propositus was homozygous for the 36.0 Kb Xba I polymorphic band and heterozygous for the Bgl II polymorphism. She was the only member of the family showing this allelic pattern. The linkage of the alleles could be determined because her mother was homozygous for the 9.0 Kb Bgl II polymorphism but heterozygous for the Xba I polymorphism. The segregation of the alleles could be traced to the proband’s son and a niece. The genotypic analysis revealed that her niece could be considered as carrying a defective gene for severe vWD.


1995 ◽  
Vol 66 (2) ◽  
pp. 109-126 ◽  
Author(s):  
Jinrui Shi ◽  
David G. Heckel ◽  
Marian R. Goldsmith

SummaryWe present data for the initial construction of a molecular linkage map for the domesticated silkworm, Bombyx mori, based on 52 progeny from an F2 cross from a pair mating of inbred strains p50 and C108, using restriction fragment length polymorphisms (RFLPs). The map contains 15 characterized single copy sequences, 36 anonymous sequences derived from a follicular cDNA library, and 10 loci corresponding to a low copy number retrotransposon, mag. The 15 linkage groups and 8 ungrouped loci account for 23 of the 28 chromosomes and span a total recombination length of 413 cM; 10 linkage groups were correlated with established classic genetic maps. Scoring data from Southern blots were analysed using two Pascal programs written specifically to analyse linkage data in Lepidoptera, where females are the heterogametic sex and have achiasmatic meiosis (no crossing-over). These first examine evidence for linkage by calculating the maximum lod score under the hypothesis that the two loci are linked over the likelihood under the hypothesis that the two loci assort independently, and then determine multilocus linkage maps for groups of putatively syntenic loci by calculating the maximum likelihood estimate of the recombination fractions and the log likelihood using the EM algorithm for a specified order of loci along the chromosome. In addition, the possibility of spurious linkage was exhaustively tested by searching for genotypes forbidden by the absence of crossing-over in one sex.


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