Development of a novel multiplexed qPCR and Pyrosequencing method for the detection of human pathogenic yersiniae

2017 ◽  
Vol 257 ◽  
pp. 247-253 ◽  
Author(s):  
M.C. Thomas ◽  
T.W. Janzen ◽  
G. Huscyzynsky ◽  
A. Mathews ◽  
K.K. Amoako
2014 ◽  
Vol 3 ◽  
Author(s):  
K. R. Kerr ◽  
S. E. Dowd ◽  
K. S. Swanson

AbstractThere has been a recent increase in the feeding of unconventional diets, including whole-prey diets, to domestic pet cats. Our objective was to characterise faecal microbial populations of domestic cats fed whole and ground (6·35 mm grind) raw 1–3-d-old chicks (Rodent Pro). Faecal samples were collected from neutered male domestic cats (mean age = 5·7 years) fed these diet items in a crossover design. Bacterial DNA was isolated from faecal samples and amplicons of the 16S rRNA V4–V6 region were generated and analysed by 454 pyrosequencing. Faecal microbial populations of cats fed whole v. ground chicks did not differ. During the study, three cats presented with symptoms of infection (anorexia or diarrhoea) and tested clinically positive for Salmonella using a standard PCR method. The remaining cats tested negative. Data were analysed post hoc to test for differences in microbial populations due to clinical status. The predominant genera were Clostridium (9–30 %), unidentified Lachnospiraceae (10–28 %), Blautia (4–19 %), Peptococcus (2–19 %) and Fusobacterium (2–14 %). Faeces of cats testing clinically positive for Salmonella had higher (P ≤ 0·05) proportions of the genera Coprococcus (5·6 v. 0·4 %) and Escherichia (subgenera Shigella; 1·1 v. 0·3 %). Salmonella was not detected in faecal samples utilising the pyrosequencing method; however, there was a shift in microbial populations due to clinical status. The clinical symptoms reported herein may be not only due to the Salmonella itself, but also shifts in other gut microbial populations.


2015 ◽  
Vol 98 (3) ◽  
pp. 563-567 ◽  
Author(s):  
Mariana Petaccia de Macêdo ◽  
Fernanda Machado de Melo ◽  
Bianca Cristina Garcia Lisboa ◽  
Louise D. Brot Andrade ◽  
Maria Dirlei Ferreira de Souza Begnami ◽  
...  

2009 ◽  
Vol 24 (S1) ◽  
pp. 1-1
Author(s):  
C.-U. Pae

Objectives:This study investigated the association between an enlarged set of SNPs at Heat shock proteins (HSPs) 70 gene and schizophrenia.Methods:Two hundred and ninety four patients with schizophrenia and 287 controls were enrolled in the study. Genotypings of 5 SNPs of HSP70 were performed using pyrosequencing method.Results:Significant association was detected at rs2075799 (allele A, Chi suare =8.03, d.f.=1 p=0.0046), but not at rs2227956 (p=0.28), rs1043618 (p=0.88), rs562047 (p=0.47) or rs539689 (p=0.32). in fact, the rs2075799*G/A genotype was more represented in patients with schizophrenia than in controls (Chi-sq=8.23, d.f.=1, p=0.0041). Haplotype based associations were also detected (global p value 0.000003); the T-A-C-C-G haplotype was more prevalent among the patients (odds ratio, OR 5.95). Sliding windows analysis revealed a major contribution from rs2227956 and rs2075799 (global-p value 0.0075), with T-A haplotype significantly associated with schizophrenia. There was no evidence of an association between the clinical variables and schizophrenia across the genotypes.Conclusion:Our results raise the possibility that HSP70 gene (i.e., haplotypes of rs2075799) might be implicated in the development of schizophrenia, although limited by rare haplotypic association with the disease. Hence further studies from different ethnics should be performed to confirm these results.


2004 ◽  
Vol 50 (12) ◽  
pp. 2392-2395 ◽  
Author(s):  
Matthew W Hruska ◽  
Reginald F Frye ◽  
Taimour Y Langaee

2014 ◽  
Vol 79 (2) ◽  
pp. 228-232 ◽  
Author(s):  
Jian R. Bao ◽  
Richard B. Clark ◽  
Ronald N. Master ◽  
Arelis E. Piscitelli ◽  
Praveena R. Tummala ◽  
...  

2015 ◽  
Vol 9 (1) ◽  
Author(s):  
Chinachote Teerapakpinyo ◽  
Phanni Wanthong ◽  
Mathawee Aumchaaumchaya ◽  
Piyamai Chankate ◽  
Warisa Kaikeaw ◽  
...  

AbstractBackground, coding for a small G-protein downstream of epidermal growth factor receptor (EGFR) plays an important role in the EGFR signaling network. Mutation inObjectivesTo develop an in-house pyrosequencing method to screen forMaterials and MethodsDNA extracted from FFPE specimens was screened forResultsOf the 74 samples with undetectable codon 12/13 mutation examined, two (2.7%) were found to harbor mutation in codon 61.ConclusionDespite the low prevalence of


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