A rapid DNA isolation procedure for the identification of Campylobacter jejuni by the polymerase chain reaction

Englen, Mark D.; Kelley, Lynda C.

doi:10.1046/j.1365-2672.2000.00841.x

Lett Appl Microbiol

2000

DOI: 10.1046/j.1365-2672.2000.00841.x

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A rapid DNA isolation procedure for the identification of Campylobacter jejuni by the polymerase chain reaction

Mark D. Englen¹,

Lynda C. Kelley²

Abstract: We have developed an efficient process for rapidly isolating campylobacter DNA using mechanical disruption combined with the guanidine‐based reagent DNAzol. Template DNA was isolated by this method from cultures of Campylobacter jejuni resistant to lysis by boiling or enzymes and identified following polymerase chain reaction (PCR) amplification using primers specific for the hippuricase gene. Direct detection of campylobacters in poultry‐processing samples by PCR is demonstrated in chicken carcass rinses spik… Show more

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Cited by 134 publications

(96 citation statements)

References 22 publications

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“…Strain CL-S1 T grew well on MA or on saline blood agar [SBA; per litre distilled water: 40 g blood agar base (BBL), 50 ml sheep blood, 30 g NaCl] at 37 u C under either aerobic or microaerobic conditions. The novel strain was preserved in marine broth 2216 (MB; Difco) supplemented with 30 % (v/v) glycerol at 280 u C.For 16S rRNA gene amplification by PCR, DNA was extracted from a single colony by a boiling method (Englen & Kelley, 2000). The crude extracts served as the DNA template for PCRs, which included Taq DNA polymerase (Bioneer) and primers 27F and 1492R (Lane, 1991).…”

mentioning

confidence: 99%

Arcobacter marinus sp. nov.

Kim

Hwang

Cho

2010

International Journal of Systematic and Evolutionary Microbiology

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A slightly curved, rod-shaped marine bacterium, designated strain CL-S1 T , was isolated from near Dokdo, an island in the East Sea, Korea. Cells were Gram-negative and grew well under either aerobic or microaerobic conditions. Analyses of the 16S rRNA and gyrA gene sequences of strain CL-S1 T revealed an affiliation with the genus Arcobacter within the class Epsilonproteobacteria. Phylogenetic analyses based on 16S rRNA and gyrA gene sequences showed that strain CL-S1 T formed a robust clade with Arcobacter halophilus LA31B T , with sequence similarities of 96.1 and 88.2 %, respectively. DNA-DNA relatedness between strain CL-S1 T and A. halophilus DSM 18005 T was 44 %, indicating that they represent genomically distinct species. Strain CL-S1 T grew optimally at 30-37 6C, at pH 7 and in the presence of 3-5 % NaCl. The dominant cellular fatty acids were iso-C 15 : 0 2-OH and/or C 16 : 1 v7c (28.4 %), C 16 : 0 (26.2 %) and C 18 : 1 v7c (22.3 %). The DNA G+C content of strain CL-S1 T was 28 mol%. Strain CL-S1 T differed phenotypically from A. halophilus LA31B T based on its ability to grow aerobically at 10 6C and inability to grow under anaerobic conditions. Based on the data presented, strain CL-S1 T is considered to represent a novel species of the genus Arcobacter, for which the name Arcobacter marinus sp. nov. is proposed. The type strain is CL-S1 T (5KCCM 90072 T 5JCM 15502 T ).The genus Arcobacter belongs to the family Campylobacteraceae, which also contains the genera Campylobacter and Sulfurospirillum ( (Neill et al., 1985;Kiehlbauch et al., 1991;Vandamme et al., 1992) and are recognized as potential emerging human pathogens (Mansfield & Forsythe, 2000). A. nitrofigilis was isolated from Spartina alterniflora roots in a salt marsh (McClung et al., 1983). A. halophilus and A. cibarius were isolated from a hypersaline lagoon (Donachie et al., 2005) and the skin of a broiler carcass (Houf et al., 2005), respectively. In the present study, a novel strain affiliated with the genus Arcobacter was isolated and subjected to a polyphasic taxonomic analysis.Seaweeds and a starfish were collected and added to a surface seawater sample (50 ml) taken from the vicinity of Dokdo, an island in the East Sea, Korea. The mixture was maintained at 4 u C for 15 days. The mixture was then homogenized by using a blender and an aliquot (50 ml) was spread onto MY medium (Bouchotroch et al., 2001). The culture was incubated aerobically at 30 u C for 2 weeks. Strain CL-S1 T was isolated and subsequently purified on fresh marine agar 2216 (MA; Difco). Strain CL-S1 T grew well on MA or on saline blood agar [SBA; per litre distilled water: 40 g blood agar base (BBL), 50 ml sheep blood, 30 g NaCl] at 37 u C under either aerobic or microaerobic conditions. The novel strain was preserved in marine broth 2216 (MB; Difco) supplemented with 30 % (v/v) glycerol at 280 u C.For 16S rRNA gene amplification by PCR, DNA was extracted from a single colony by a boiling method (Englen & Kelley, 2000). The crude extracts served as the DNA template for...

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mentioning

confidence: 99%

Arcobacter marinus sp. nov.

Kim

Hwang

Cho

2010

International Journal of Systematic and Evolutionary Microbiology

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“…For 16S rRNA gene amplification by PCR, DNA was extracted from a single colony by a boiling method (Englen & Kelley, 2000). The crude extracts served as the DNA template for PCR, which included Taq DNA polymerase (Bioneer) and primers 27F and 1492R (Lane, 1991).…”

mentioning

confidence: 99%

Maribacter antarcticus sp. nov., a psychrophilic bacterium isolated from a culture of the Antarctic green alga Pyramimonas gelidicola

Zhang¹,

Hwang²,

Kang³

et al. 2009

INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLO

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“…For 16S rRNA gene amplification by PCR, DNA was extracted from a single colony based on a boiling method (Englen & Kelley, 2000). The crude extracts served as the DNA template for PCRs, which included Taq DNA polymerase (Bioneer) and primers 27F and 1492R (Lane, 1991).…”

mentioning

confidence: 99%

Pelagibius litoralis gen. nov., sp. nov., a marine bacterium in the family Rhodospirillaceae isolated from coastal seawater

Choi

Hwang

Cho

2009

INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLO

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Pelagibius litoralis gen. nov., sp. nov., a marine bacterium in the family Rhodospirillaceae isolated from coastal seawater A Gram-negative, strictly aerobic, slightly curved rod-shaped bacterial strain, designated CL-UU02 T , was isolated from coastal seawater off the east coast of Korea. 16S rRNA gene sequence analysis revealed a clear affiliation of this novel strain with the family Rhodospirillaceae. Strain CL-UU02 T formed a robust cluster with the type strains of species of the genus Rhodovibrio at 16S rRNA gene sequence similarity levels of 89.9-90.4 %. Strain CL-UU02 T shared no more than 89 % 16S rRNA gene sequence similarity with the type strains of other species in the family Rhodospirillaceae. Strain CL-UU02 T was able to grow in the presence of 2-6 % sea salts, and grew optimally at 28-30 6C and pH 7-8. The DNA G+C content of strain CL-UU02 T was 66.3 mol%. On the basis of phylogenetic analyses and chemotaxonomic and physiological data, strain CL-UU02 T is considered to represent a novel species of a new genus in the family Rhodospirillaceae, for which the name Pelagibius litoralis gen. nov., sp. nov. is proposed. The type strain of Pelagibius litoralis is CL-UU02 T (5KCCM 42323 T 5JCM 15426 T ).

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A rapid DNA isolation procedure for the identification of Campylobacter jejuni by the polymerase chain reaction

Cited by 134 publications

References 22 publications

Arcobacter marinus sp. nov.

Arcobacter marinus sp. nov.

Maribacter antarcticus sp. nov., a psychrophilic bacterium isolated from a culture of the Antarctic green alga Pyramimonas gelidicola

Pelagibius litoralis gen. nov., sp. nov., a marine bacterium in the family Rhodospirillaceae isolated from coastal seawater

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