Species-Specific Plasmid Sequences for PCR Identification of the Three Species of Borrelia burgdorferiSensu Lato  Involved in Lyme Disease

Species-specific sequences were shown to be carried by plasmids of the three main species of Borrelia burgdorferi sensu lato involved in Lyme disease. Libraries of the 16-, 33-, and 25-kb plasmids of B. burgdorferi sensu stricto, Borrelia garinii, and Borrelia afzelii, respectively, were then built and used to isolate species-specific sequences. After sequencing of the cloned inserts, three sets of primers were designed. They were shown to determine species-specific PCR amplification products. The sensitivities of the PCR assay with these primers were 100 spirochetes for B. burgdorferi sensu stricto and 1,000 spirochetes for B. garinii and B. afzelii. The usefulness of these primers for the identification of species in biological samples (tick, serum, and cerebrospinal fluid samples) was ascertained.

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Specific PCR Detection of Arcobacter butzleri, Arcobacter cryaerophilus, Arcobacter skirrowii, and Arcobacter cibarius in Chicken Meat

Journal of Food Protection ◽

10.4315/0362-028x-72.7.1491 ◽

2009 ◽

Vol 72 (7) ◽

pp. 1491-1495 ◽

Cited By ~ 16

Author(s):

DANIELA PENTIMALLI ◽

NICOLETTE PEGELS ◽

TERESA GARCÍA ◽

ROSARIO MARTÍN ◽

ISABEL GONZÁLEZ

Keyword(s):

Pcr Amplification ◽

Chicken Meat ◽

Pcr Analysis ◽

Rrna Gene ◽

Pcr Assay ◽

Gyra Gene ◽

Specific Primers ◽

Specific Pcr ◽

Arcobacter Butzleri ◽

Species Specific

An enrichment PCR assay using species-specific primers was developed for the detection of Arcobacter butzleri, Arcobacter cryaerophilus, Arcobacter skirrowii, and Arcobacter cibarius in chicken meat. Primers for A. cryaerophilus, A. skirrowii, and A. cibarius were designed based on the gyrA gene to amplify nucleic acid fragments of 212, 257, and 145 bp, respectively. The A. butzleri–specific primers were designed flanking a 203-bp DNA fragment in the 16S rRNA gene. The specificity of the four primer pairs was assessed by PCR analysis of DNA from a panel of Arcobacter species, related Campylobacter, Helicobacter species, and other food bacteria. The applicability of the method was then validated by testing 42 fresh retail-purchased chicken samples in the PCR assay. An 18-h selective preenrichment step followed by PCR amplification with the four Arcobacter primer sets revealed the presence of Arcobacter spp. in 85.7% of the retail chicken samples analyzed. A. butzleri was the only species present in 50% of the samples, and 35.7% of the samples were positive for both A. butzleri and A. cryaerophilus. A. skirrowii and A. cibarius were not detected in any of the chicken samples analyzed. The enrichment PCR assay developed is a specific and rapid alternative for the survey of Arcobacter contamination in meat.

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Species-specific PCR assay for L. infantum/L. donovani discrimination

Acta Tropica ◽

10.1016/j.actatropica.2006.10.012 ◽

2006 ◽

Vol 100 (3) ◽

pp. 241-245 ◽

Cited By ~ 36

Author(s):

Mallorie Hide ◽

Anne-Laure Bañuls

Keyword(s):

Pcr Assay ◽

Specific Pcr ◽

Specific Pcr Assay ◽

Species Specific

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A SPECIES-SPECIFIC PCR ASSAY BASED ON THE INTERNAL TRANSCRIBED SPACER (ITS) REGIONS FOR IDENTIFICATION OF MYCOSPHAERELLA EUMUSAE, M. FIJIENSIS AND M. MUSICOLA ON BANANA

BIOTROPIA ◽

10.11598/btb.2012.19.1.232 ◽

2012 ◽

Vol 19 (1) ◽

Keyword(s):

Internal Transcribed Spacer ◽

Pcr Assay ◽

Specific Pcr ◽

Its Regions ◽

Specific Pcr Assay ◽

Species Specific

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DNA Microarray Assessment of Putative Borrelia burgdorferi Lipoprotein Genes

Infection and Immunity ◽

10.1128/iai.70.6.3300-3303.2002 ◽

2002 ◽

Vol 70 (6) ◽

pp. 3300-3303 ◽

Cited By ~ 30

Author(s):

Fang Ting Liang ◽

F. Kenneth Nelson ◽

Erol Fikrig

Keyword(s):

Lyme Disease ◽

Borrelia Burgdorferi ◽

Dna Microarray ◽

Sensu Stricto ◽

Borrelia Garinii ◽

Borrelia Afzelii ◽

Genes Encoding ◽

The Individual

ABSTRACT A DNA microarray containing fragments of 137 Borrelia burgdorferi B31 putative lipoprotein genes was used to examine Lyme disease spirochetes. DNA from B. burgdorferi sensu stricto B31, 297, and N40; Borrelia garinii IP90; and Borrelia afzelii P/Gau was fluorescently labeled and hybridized to the microarray, demonstrating the degree to which the individual putative lipoprotein genes were conserved among the genospecies. These data show that a DNA microarray can globally examine the genes encoding B. burgdorferi lipoproteins.

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Far-Reaching Dispersal of Borrelia burgdorferi Sensu Lato-Infected Blacklegged Ticks by Migratory Songbirds in Canada

Healthcare ◽

10.3390/healthcare6030089 ◽

2018 ◽

Vol 6 (3) ◽

pp. 89 ◽

Cited By ~ 8

Author(s):

John Scott ◽

Kerry Clark ◽

Janet Foley ◽

Bradley Bierman ◽

Lance Durden

Keyword(s):

Lyme Disease ◽

Borrelia Burgdorferi ◽

Health Care Providers ◽

Ixodes Scapularis ◽

Pcr Amplification ◽

Sensu Stricto ◽

Borrelia Burgdorferi Sensu Lato ◽

Care Providers ◽

Northern Areas ◽

Migratory Songbirds

Lyme disease has been documented in northern areas of Canada, but the source of the etiological bacterium, Borrelia burgdorferi sensu lato (Bbsl) has been in doubt. We collected 87 ticks from 44 songbirds during 2017, and 24 (39%) of 62 nymphs of the blacklegged tick, Ixodes scapularis, were positive for Bbsl. We provide the first report of Bbsl-infected, songbird-transported I. scapularis in Cape Breton, Nova Scotia; Newfoundland and Labrador; north-central Manitoba, and Alberta. Notably, we report the northernmost account of Bbsl-infected ticks parasitizing a bird in Canada. DNA extraction, PCR amplification, and DNA sequencing reveal that these Bbsl amplicons belong to Borrelia burgdorferi sensu stricto (Bbss), which is pathogenic to humans. Based on our findings, health-care providers should be aware that migratory songbirds widely disperse B. burgdorferi-infected I. scapularis in Canada’s North, and local residents do not have to visit an endemic area to contract Lyme disease.

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Species-Specific and Ubiquitous-DNA-Based Assays for Rapid Identification of Staphylococcus aureus

Journal of Clinical Microbiology ◽

10.1128/jcm.36.3.618-623.1998 ◽

1998 ◽

Vol 36 (3) ◽

pp. 618-623 ◽

Cited By ~ 215

Author(s):

Francis Martineau ◽

François J. Picard ◽

Paul H. Roy ◽

Marc Ouellette ◽

Michel G. Bergeron

Keyword(s):

Staphylococcus Aureus ◽

Genomic Library ◽

Antibiotic Resistance Genes ◽

Pcr Amplification ◽

Rapid Identification ◽

Clinical Microbiology Laboratory ◽

Chromosomal Dna ◽

Pcr Assay ◽

Serious Infections ◽

Species Specific

Staphylococcus aureus is the cause of serious infections in humans, including endocarditis, deep-seated abscesses, and bacteremia, which lead to toxic and septic shock syndromes. Rapid and direct identification of this bacterium specifically and ubiquitously directly from clinical specimens would be useful in improving the diagnosis of S. aureus infections in the clinical microbiology laboratory. A wide variety of kits based on biochemical characteristics efficiently identify S. aureus, but the rapidity and the accuracy of each of these methods combined with testing of clinically relevant antibiotic resistance genes need to be improved. On the basis of hybridization assays with randomly selected clones from an S. aureus genomic library, we have identified a chromosomal DNA fragment which is specific for S. aureus and which detected all 82 S. aureus isolates tested. This 442-bp fragment was sequenced and was used to design a set of PCR amplification primers. The PCR assay was also specific and ubiquitous for the identification from bacterial cultures of 195 clinical strains of S. aureus isolated from a variety of anatomical sites and obtained from hospitals throughout the world. The PCR assay that we have developed is simple and can be performed in about 1 h. This DNA-based test provides a novel diagnostic tool for the diagnosis of S. aureus infections.

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Rate of Infection of Ixodes ricinus Ticks with Borrelia burgdorferi Sensu Stricto, Borrelia garinii, Borrelia afzelii and Group VS116 in an Endemic Focus of Lyme Disease in Italy

European Journal of Clinical Microbiology & Infectious Diseases ◽

10.1007/s100960050023 ◽

1998 ◽

Vol 17 (2) ◽

pp. 90-94 ◽

Cited By ~ 5

Author(s):

M. Cinco ◽

D. Padovan ◽

R. Murgia ◽

L. Poldini ◽

L. Frusteri ◽

...

Keyword(s):

Lyme Disease ◽

Borrelia Burgdorferi ◽

Ixodes Ricinus ◽

Sensu Stricto ◽

Borrelia Garinii ◽

Borrelia Afzelii ◽

Endemic Focus ◽

Ixodes Ricinus Ticks ◽

Borrelia Burgdorferi Sensu Stricto

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A simple method for discriminating Bursaphelenchus xylophilus and B. mucronatus by species-specific polymerase chain reaction primer pairs

Nematology ◽

10.1163/1568541041217960 ◽

2004 ◽

Vol 6 (2) ◽

pp. 273-277 ◽

Cited By ~ 38

Author(s):

Koji Matsunaga ◽

Katsumi Togashi

Keyword(s):

Pcr Amplification ◽

Nematode Species ◽

Bursaphelenchus Xylophilus ◽

Polymerase Chain Reaction Primer ◽

Dna Fragments ◽

Simple Method ◽

Specific Pcr ◽

Pcr Products ◽

Specific Pcr Primer ◽

Species Specific

Abstract Two species-specific PCR primer pairs were developed for identifying the two nematode species, Bursaphelenchus xylophilus and B. mucronatus. The primer pairs were developed from the sequence of ribosomal DNA (rDNA) repeats to produce DNA fragments of different lengths by PCR amplification. The DNA fragments for B. mucronatus and B. xylophilus were 210 bp and 557 bp, respectively. When mixed, neither primer pair inhibited the PCR amplification of the other. Five isolates of B. xylophilus and four isolates of B. mucronatus showed different band profiles of PCR products between the two species, but identical profiles among isolates of the same species.

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