scholarly journals Genetic Profiling of Aggregatibacter actinomycetemcomitans Serotype B Isolated from Periodontitis Patients Living in Sweden

Pathogens ◽  
2019 ◽  
Vol 8 (3) ◽  
pp. 153 ◽  
Author(s):  
Anders Johansson ◽  
Rolf Claesson ◽  
Carola Höglund Åberg ◽  
Dorte Haubek ◽  
Mark Lindholm ◽  
...  
Keyword(s):  
Diagnostic Marker ◽  
Aggregatibacter Actinomycetemcomitans ◽  
Conventional Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Genetic Profiling ◽  
Attachment Loss ◽  
Polymerase Chain ◽  
Serotype B ◽  
Periodontal Attachment Loss ◽  
Potential Use

The bacterium Aggregatibacter actinomycetemcomitans is associated with aggressive forms of periodontitis and with systemic diseases, such as endocarditis. By assessing a Ghanaian longitudinal adolescent cohort, we earlier recognized the cagE gene as a possible diagnostic marker for a subgroup of JP2 and non-JP2 genotype serotype b A. actinomycetemcomitans strains, associated with high leukotoxicity as determined in a semi-quantitative cell assay. This group of A. actinomycetemcomitans is associated with the progression of attachment loss. In the present work, we used conventional polymerase chain reaction (PCR) and quantitative PCR to perform the cagE genotyping of our collection of 116 selected serotype b A. actinomycetemcomitans strains, collected over a period of 15 years from periodontitis patients living in Sweden. The A. actinomycetemcomitans strains carrying cagE (referred to as cagE+; n = 49) were compared to the cagE-negative strains (n = 67), present at larger proportions in the subgingival plaque samples, and were also much more prevalent in the young (≤35 years) compared to in the old (>35 years) group of patients. Our present results underline the potential use of cagE genotyping in the risk assessment of the development of periodontal attachment loss in Swedish adolescents.

scholarly journals Detection of MYCN Amplification in Serum DNA Using Conventional Polymerase Chain Reaction

2016 ◽  
Vol 31 (9) ◽  
pp. 1392 ◽  
Author(s):  
Youngeun Ma ◽  
Ji Won Lee ◽  
Soo Jin Park ◽  
Eun Sang Yi ◽  
Young Bae Choi ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Conventional Polymerase Chain Reaction ◽  
Mycn Amplification ◽  
Chain Reaction ◽  
Serum Dna ◽  
Polymerase Chain

scholarly journals Assessment of Aggregatibacter actinomycetemcomitans and Prevotella intermedia Counts around Healthy Implants, Diseased Implants and Sound Teeth: A Preliminary Study

2017 ◽  
Vol 9 (1) ◽  
pp. 18-25 ◽  
Author(s):  
Siamak Yaghobee ◽  
Afshin Khorsand ◽  
Nojan Jahedmanesh ◽  
Mahdi Kadkhodazadeh
Keyword(s):  
Polymerase Chain Reaction ◽  
Gingival Crevicular Fluid ◽  
Aggregatibacter Actinomycetemcomitans ◽  
Prevotella Intermedia ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Standard Curve ◽  
Preliminary Study ◽  
Polymerase Chain ◽  
Crevicular Fluid

Background. This study aimed to assess Aggregatibacter actinomycetemcomitans (Aa) and Prevotella intermedia (PI) counts in gingival crevicular fluid (GCF) around healthy implants, diseased implants and sound teeth. Methods. Eight patients (four males and four females), who had healthy implants, implants with peri-implantitis and sound teeth, were selected. Samples (GCF) were analyzed using real-time polymerase chain reaction (RT-PCR). The above-mentioned bacteria were detected and counted. Data analysis in RT-PCR was carried out based on the standard curve using Prism software to compare Pi and Aa counts between the three areas (GCF around sound teeth, healthy implants and implants with peri-implantitis). Results. Pi counts were significantly higher in GCF around implants with peri-implantitis (8 implants) than around healthy implants (8 implants) (P<0.001) and sound teeth (8) (P=0.012). No significant differences were found in Pi counts in GCF around healthy implants and sound teeth (P=0.063). Aa counts in GCF around implants with peri-implantitis were significantly higher than those around healthy implants (P=0.002) and sound teeth (P=0.024). No significant differences were noted in Aa counts in GCF around healthy implants and sound teeth (P=0.57). Conclusion. Aa and Pi counts in GCF around diseased implants were higher than around healthy implants and sound teeth. Also, Aa counts were significantly higher than Pi counts.

Seizures as a Consequence of Hyperviscosity Syndrome in Two Dogs Naturally Infected with Leishmania infantum

2016 ◽  
Vol 52 (2) ◽  
pp. 119-123 ◽  
Author(s):  
Daniela Proverbio ◽  
Eva Spada ◽  
Roberta Perego ◽  
Giada Bagnagatti de Giorgi
Keyword(s):  
Monoclonal Gammopathy ◽  
Conventional Polymerase Chain Reaction ◽  
Immunofluorescence Assay ◽  
Canine Leishmaniasis ◽  
Chain Reaction ◽  
Short Term ◽  
Hyperviscosity Syndrome ◽  
Clinical Presentations ◽  
Polymerase Chain ◽  
Clinical Complaint

Serum hyperviscosity syndrome (HVS) was documented in two dogs with canine leishmaniasis (CanL) and seizures as the major clinical complaint. In both cases, laboratory abnormalities included mild non-regenerative anemia, thrombocytopenia, hypoalbuminemia, hyperproteinemia with monoclonal gammopathy, and marked serum hyperviscosity. CanL was diagnosed using cytology in one case and indirect immunofluorescence assay and conventional polymerase chain reaction in the second. Specific therapy with meglumine antimoniate and allopurinolc led to short-term remission in both dogs and normalization of serum viscosity. Although dogs rarely develop HVS, it should be suspected if hyperproteinemia and monoclonal gammopathy are present. Since CanL manifests with a variety of clinical presentations, including seizures resulting from HVS-induced central nervous system hypoxia, it should also be considered as a differential diagnosis in animals with seizures as a primary presenting sign.

scholarly journals A Comparative Analysis of Polymerase Chain Reaction and Direct Fluorescent Antibody Test for Diagnosis of Genital Herpes

2017 ◽  
Vol 9 (01) ◽  
pp. 053-056 ◽  
Author(s):  
Vrushali Patwardhan ◽  
Preena Bhalla ◽  
Deepti Rawat ◽  
Vijay Kumar Garg ◽  
Kabir Sardana ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Genital Herpes ◽  
Fluorescent Antibody ◽  
Antibody Test ◽  
Conventional Polymerase Chain Reaction ◽  
Genital Ulcer ◽  
Chain Reaction ◽  
Direct Fluorescent Antibody ◽  
Polymerase Chain ◽  
Simplex Virus

ABSTRACT Objective: To compare laboratory tests that can simultaneously detect and type herpes simplex virus (HSV) directly from the genital ulcer specimens in clinically suspected cases of genital herpes. Materials and Methods: A study was conducted over 10 months and 44 adult male and female patients clinically suspected with genital herpes were recruited. Genital ulcer swab specimens were subjected to glycoprotein-G gene-based conventional polymerase chain reaction (PCR) and commercially available direct fluorescent antibody (DFA) test and the results were compared. Results: PCR for HSV was positive in 82% (36/44) cases. DFA was positive in 68.2% (30/44) cases. There was 100% agreement between HSV types detected by DFA and PCR. The strength of agreement between the results was better in primary genital herpes than recurrent cases. Conclusion: PCR was found to be better in the detection of HSV in recurrent genital herpes patients. It is a better modality, especially when genital herpes clinically presents with ulcerative or crusted lesions, and is also a cheaper alternative as compared to DFA.

scholarly journals First detection of Pasteurella multocida type B:2 in Hungary associated with systemic pasteurellosis in backyard pigs

2015 ◽  
Vol 63 (2) ◽  
pp. 141-156 ◽  
Author(s):  
Barbara Ujvári ◽  
Levente Szeredi ◽  
László Pertl ◽  
Gergely Tóth ◽  
Károly Erdélyi ◽  
...  
Keyword(s):  
Small Area ◽  
Pasteurella Multocida ◽  
Sequence Type ◽  
Type B ◽  
Chain Reaction ◽  
Haemorrhagic Septicaemia ◽  
Gene Typing ◽  
Polymerase Chain ◽  
Serotype B ◽  
Backyard Pigs

This is the first report of Pasteurella multocida type B in Hungarian pigs. This disease was observed in backyard-raised pigs in three households within a small area. Neither the source of the infection nor the epidemiological connection between any of the premises could be determined. The most consistent lesion was dark red discolouration of the skin of the ventral neck and brisket, with accompanying oedema and haemorrhages. The morbidity was low and lethality relatively high, with three dead (50%) and two euthanised (33%) out of six affected animals. A total of three isolates of P. multocida (P55, P56 and P57) were cultured from these cases and examined in detail. These were identified as P. multocida ssp. multocida biovar 3. All were toxA negative and belonged to serotype B:2. Multilocus sequence typing was used to assign these to a new sequence type (ST61) that is closely related to other haemorrhagic septicaemia causing strains of P. multocida regardless of the host. M13 polymerase chain reaction and virulence-associated gene typing also show that type B strains form a highly homogeneous, distinct phylogenic group within P. multocida.

scholarly journals Discrimination between Mycoplasma and Acholeplasma species of bovine origin using digitonin disc diffusion assay, nisin disc diffusion assay, and conventional polymerase chain reaction

2011 ◽  
Vol 24 (1) ◽  
pp. 7-13 ◽  
Author(s):  
Sukolrat Boonyayatra ◽  
Lawrence K. Fox ◽  
John M. Gay ◽  
Ashish Sawant ◽  
Thomas E. Besser
Keyword(s):  
Polymerase Chain Reaction ◽  
Growth Inhibition ◽  
Gold Standard ◽  
Conventional Polymerase Chain Reaction ◽  
Disc Diffusion ◽  
Chain Reaction ◽  
Disc Diffusion Assay ◽  
Field Isolates ◽  
Polymerase Chain ◽  
Diffusion Assay

Microbiological culture of milk samples has been used as a standard diagnosis for Mycoplasma mastitis. This technique is effective in isolating mollicutes that are Mycoplasma-like; however, isolates may be misinterpreted as Acholeplasma species, which are indistinguishable from Mycoplasma species by culture. A study to contrast the abilities of 2 culture-based tests, digitonin and nisin disc diffusion assays and a conventional polymerase chain reaction (PCR) technique, to discriminate between Mycoplasma and Acholeplasma was performed using 16S ribosomal RNA gene partial sequencing as the gold standard of comparison. A total of 288 bovine mollicute field isolates (248 from milk and 40 from other organ sites) and 13 reference strains were tested. Results obtained from the digitonin disc diffusion assay when it was performed with all field isolates were 92.7% and 99.0% in agreement with the gold standard using 5 mm and 3 mm of zone of growth inhibition as thresholds, respectively. Considering only milk isolates, agreements between the digitonin disc diffusion assay with the gold standard were 97.2% and 100% using 5 mm and 3 mm of zone of growth inhibition as thresholds, respectively. Culture identification using the nisin disc diffusion assay and the PCR was in a 100% agreement with the gold standard. Comparable results using culture-based nisin and digitonin disc diffusion assays, and PCR, to distinguish Mycoplasma and Acholeplasma species was found, especially for isolates from bovine milk.

scholarly journals A Two-Step Approach for Diagnosing Glutamate Dehydrogenase Genes by Conventional Polymerase Chain Reaction from Clostridium difficile Isolates

2019 ◽  
Vol 11 (3) ◽  
pp. 135-140
Author(s):  
Sepideh Khodaparast ◽  
Ashraf Mohabati Mobarez ◽  
Mehdi Saberifiroozi
Keyword(s):  
Polymerase Chain Reaction ◽  
Clostridium Difficile ◽  
Glutamate Dehydrogenase ◽  
Conventional Polymerase Chain Reaction ◽  
Conventional Pcr ◽  
Accessory Gene ◽  
Chain Reaction ◽  
Fecal Samples ◽  
Genotype 2 ◽  
Polymerase Chain

BACKGROUND Clostridium difficile is the major causative agent of nosocomial antibiotic-associated colitis. The gold standard for C. difficile detection is stool culture followed by cytotoxic assay, although it is laborious and time-consuming. We developed a screening test based on a two-step conventional polymerase chain reaction (PCR) approach to detect gluD, the glutamate dehydrogenase (GDH) enzyme gene, which is a marker for screening of C. difficile. Targeting gluD comparing to the conserved stable genetic element of pathogenicity locus (PaLoc), with an accessory gene of Cdd3, was an effective method for the detection of this pathogen from patients with enterocolitis. METHODS Fresh fecal samples of the patients who were clinically suspicious for antibiotic-associated colitis were collected. Stool specimens were cultured on the cycloserine-cefoxitin fructose agar (CCFA) in an anaerobic condition, following alcohol shock treatment and enrichment in Clostridium difficile Brucella broth (CDBB). On confirmed colonies, PCR was carried out for detection of PaLoc subsidiary gene, Cdd3, and toxicogenic genes, tcdA and tcdB. The gluD that is GDH gene detection was performed by conventional PCR on the extracted DNA from 578 fresh stool samples. RESULTS 57 (9.8%) strains of C. difficile were approved by conventional PCR for gluD and Cdd3 genes, in which 37 (6.4%) colonies had tcdA+/tcdB+ genotype, 2 (0.3%) tcdA+/tcdB-, 4 (0.7%) tcdA-/ tcdB+ and the remaining 14 (2.4%) colonies were tcdA and tcdB negative. CONCLUSION These results demonstrate that targeting gluD by PCR is quite promising for rapid detection of C. difficile from fresh fecal samples. Furthermore, the multiple-gene analysis for tcdA and tcdB assay proved a reliable approach for diagnosing of toxigenic strains among clinical samples.

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