Identification of streptococci isolated from various sources by determination ofcfbgene and other CAMP-factor genes

In the present study, the CAMP-factor (cfb) gene of streptococci of serological group B (Streptococcus agalactiae) and the CAMP-factor (cfu) gene of S. uberis could be amplified by polymerase chain reaction. A cfb specific amplicon could be observed for all 128 phenotypically CAMP-positive S. agalactiae, for the phenotypically CAMP-negative S. agalactiae strain 74-360, and for 2 S. difficile reference strains. A cfu specific amplicon could be observed for all 7 phenotypically CAMP-positive S. uberis. Four S. agalactiae strains isolated from 4 cows with mastitis appeared to be phenotypically CAMP-negative and negative in the cfb gene PCR. The CAMP-positive and CAMP-negative isolates, including both S. difficile, could be identified as S. agalactiae by amplification of a S. agalactiae specific part of the V2 region of the 16S rRNA and a species-specific part of the 16S-23S rRNA intergenic spacer region. Amplification of an internal fragment of the cfb gene with a reduced annealing temperature yielded positive reactions not only for CAMP-positive S. agalactiae, but also for phenotypically CAMP-positive S. pyogenes (n = 4), S. canis (n = 28), and S. uberis (n = 7), indicating a close relation of the CAMP genes of these 4 species. The relation could be further demonstrated by sequencing the internal fragment of the CAMP-factor (cfg) gene of S. canis and comparing the sequence with those of S. agalactiae, S. pyogenes, and S. uberis.Key words: CAMP factor, cfb, cfu, S. canis.

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Genomic fingerprinting ofFrankiamicrosymbionts fromCeanothuscopopulations using repetitive sequences and polymerase chain reactions

Canadian Journal of Botany ◽

10.1139/b99-069 ◽

1999 ◽

Vol 77 (9) ◽

pp. 1220-1230 ◽

Cited By ~ 3

Author(s):

Soon-Chun Jeong ◽

David D Myrold

Keyword(s):

Dna Sequencing ◽

Dna Analysis ◽

Repetitive Sequences ◽

Intergenic Spacer ◽

Rrna Genes ◽

23S Rrna ◽

Genomic Fingerprinting ◽

Chain Reactions ◽

Intergenic Spacer Region ◽

Polymerase Chain

Specificity between Ceanothus species and their microsymbionts, Frankia, were investigated with nodules collected from three geographically separated copopulations of Ceanothus species. Nodules were analyzed using DNA sequencing and repetitive sequence polymerase chain reaction (rep-PCR) techniques. DNA sequencing of the intergenic spacer region between 16S and 23S rRNA genes suggested that Ceanothus-microsymbiotic Frankia are closely related at the intraspecific level. Diversity of the microsymbionts was further analyzed by genomic fingerprinting using repetitive sequences and PCR. A newly designed direct repeat (DR) sequence and a BOX sequence were used as PCR primers after justification that these primers can generate Frankia-specific fingerprints from nodule DNA. Analysis of the nodules using BOX- and DR-PCR showed that Ceanothus-microsymbiotic Frankia exhibited less diversity within each copopulation than among copopulations. These data suggested that geographic separation plays a more important role for divergence of Ceanothus-microsymbiotic Frankia than host plant.Key words: Frankia, Ceanothus, rep-PCR, diversity.

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Molecular Detection and Characterization of Borrelia garinii (Spirochaetales: Borreliaceae) in Ixodes nipponensis (Ixodida: Ixodidae) Parasitizing a Dog in Korea

Pathogens ◽

10.3390/pathogens8040289 ◽

2019 ◽

Vol 8 (4) ◽

pp. 289 ◽

Cited By ~ 2

Author(s):

Seung-Hun Lee ◽

Youn-Kyoung Goo ◽

Paul John L. Geraldino ◽

Oh-Deog Kwon ◽

Dongmi Kwak

Keyword(s):

Molecular Detection ◽

Single Gene ◽

Protein A ◽

Intergenic Spacer ◽

Surface Protein ◽

23S Rrna ◽

Borrelia Garinii ◽

Intergenic Spacer Region ◽

Individual Level ◽

Pcr Targeting

The present study aimed to detect and characterize Borrelia spp. in ticks attached to dogs in Korea. Overall, 562 ticks (276 pools) attached to dogs were collected and tested for Borrelia infection by PCR targeting the 5S-23S rRNA intergenic spacer region (rrf-rrl). One tick larva (pool level, 0.4%; individual level, 0.2%) was confirmed by sequencing Borrelia garinii, a zoonotic pathogen. For molecular characterization, the outer surface protein A (ospA) and flagellin genes were analyzed. Phylogenetic ospA analysis distinguished B. garinii from B. bavariensis, which has been recently identified as a novel Borrelia species. On the other hand, phylogenetic analysis showed that single gene analysis involving rrf-rrl or flagellin was not sufficient to differentiate B. garinii from B. bavariensis. In addition, the B. garinii-infected tick was identified as Ixodes nipponensis by sequencing according to mitochondrial 16S rRNA and the second transcribed spacer region. To our knowledge, this is the first study to report the molecular detection of B. garinii in I. nipponensis parasitizing a dog in Korea. Continuous monitoring of tick-borne pathogens in ticks attached to animals is required to avoid disease distribution and possible transmission to humans.

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Differentiation of group VIII Spiroplasma strains with sequences of the 16S–23S rDNA intergenic spacer region

Canadian Journal of Microbiology ◽

10.1139/w04-091 ◽

2004 ◽

Vol 50 (12) ◽

pp. 1061-1067 ◽

Cited By ~ 14

Author(s):

Laura B Regassa ◽

Kimberly M Stewart ◽

April C Murphy ◽

Frank E French ◽

Tao Lin ◽

...

Keyword(s):

Intergenic Spacer ◽

Analytical Models ◽

23S Rrna ◽

Rrna Gene ◽

Group Viii ◽

Similarity Coefficients ◽

Intergenic Spacer Region ◽

Intergenic Sequences ◽

The Stability ◽

The 16S Rrna Gene

Spiroplasma species (Mollicutes: Spiroplasmataceae) are associated with a wide variety of insects, and serology has classified this genus into 34 groups, 3 with subgroups. The 16S rRNA gene has been used for phylogenetic analysis of spiroplasmas, but this approach is uninformative for group VIII because the serologically distinct subgroups generally have similarity coefficients >0.990. Therefore, we investigated the utility of the 16S–23S rRNA spacer region as a means to differentiate closely related subgroups or strains. We generated intergenic sequences and detailed serological profiles for 8 group VIII Spiroplasma strains. Sequence analyses using Maximum Parsimony, Neighbor Joining, and Maximum Likelihood placed the strains into 2 clades. One clade consisted of strains BARC 2649 and GSU5367. The other clade was divided into clusters containing representatives of the 3 designated group VIII subgroups (EA-1, DF-1, and TAAS-1) and 3 previously unclassified strains. The stability of the positions of the strains in various analytical models and the ability to provide robust support for groupings tentatively supported by serology indicates that the 16S–23S intergenic rDNA sequence will prove useful in intragroup analysis of group VIII spiroplasmas.Key words: Mollicutes, Spiroplasma, phylogeny, Tabanidae.

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A putative transposase gene in the 16S–23S rRNA intergenic spacer region of Mycoplasma imitans

Microbiology ◽

10.1099/mic.0.26629-0 ◽

2004 ◽

Vol 150 (4) ◽

pp. 1023-1029 ◽

Cited By ~ 28

Author(s):

Ryô Harasawa ◽

David G. Pitcher ◽

Ana S. Ramírez ◽

Janet M. Bradbury

Keyword(s):

Relative Size ◽

Intergenic Spacer ◽

Its Region ◽

23S Rrna ◽

Mycoplasma Species ◽

Insertion Element ◽

Transposase Gene ◽

Intergenic Spacer Region ◽

Pcr Products ◽

Type Strains

Examination of the nucleotide sequences of the 16S–23S intergenic transcribed spacer (ITS) region of Mycoplasma imitans and Mycoplasma gallisepticum identified a putative transposase gene located only in the ITS of M. imitans, which can be used as a genetic marker to distinguish these two species. The relative size of the PCR products of the ITS region allowed a clear distinction to be made between strains of M. imitans and M. gallisepticum, both of which could be readily discriminated from the type strains of all the other recognized avian Mycoplasma species. In addition, the putative transposase gene assigned in the ITS of M. imitans was shown to include a sequence homologous to that of the P75 gene of M. gallisepticum. This is believed to be the first description of an insertion element in the rRNA operon region of a mycoplasma species.

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