scholarly journals A New Specific Sequence to Distinguish B.canis From Other Brucella by PCR

2021 ◽  
Author(s):  
YinBo YE ◽  
JiangHua YANG ◽  
DongLiang LI ◽  
LiHua HAO ◽  
Zhao ZHANG ◽  
...  
Keyword(s):  
Genome Comparison ◽  
Detection Accuracy ◽  
Comparison Analysis ◽  
Pcr Assay ◽  
Specific Primers ◽  
Specific Sequence ◽  
Specific Pcr ◽  
Pcr Method ◽  
Specific Pcr Assay ◽  
Canine Brucellosis

Abstract Background: Brucellosis is a zoonotic disease worldwide. The increasing number of pet dogs has raised new risk of people getting canine brucella with absent or mild symptom. Besides, the canine brucellosis can be caused by other brucella species, so their infection could be omitted by the PCR method. The present PCR methods can only detect canine brucella, by which cases infected with other brucella would appear negative. It’s an urge to develop a specific PCR assay for detecting canine Brucellosis, Whether the pathogen was B.canis or any other Brucella.Resaults: a differential sequence of B.canis were found by genome comparison analysis and were analyzed by BLAST. Then a PCR method was established using specific primers in the sequence and tested for clinical application. It could detect canine brucellosis caused by B.canis or other Brucella species with 310-bp and 413-bp product, respectively. The developed PCR method had specificity for non-brucella and a sensitivity of 100 copies of Brucella DNA. The detection accuracy verified with spiked samples was 95.5% (21/22) for B.canis and 100% (22/22) for other brucella. Conclusions: The study found a specific sequence of B.canis and developed a PCR detection method to detect canine brucellosis caused by B.canis or other Brucella species. The method established in this study will more comprehensively detect the pathogen of canine brucellosis and provide important methods and means for preventing and controlling this disease.

A new Brucella canis species-specific PCR assay for the diagnosis of canine brucellosis

2014 ◽  
Vol 37 (4) ◽  
pp. 237-241 ◽  
Author(s):  
Sung-Il Kang ◽  
Sang-Eun Lee ◽  
Ji-Yeon Kim ◽  
Kichan Lee ◽  
Jong-Wan Kim ◽  
...  
Keyword(s):  
Pcr Assay ◽  
Specific Pcr ◽  
Brucella Canis ◽  
Specific Pcr Assay ◽  
Species Specific ◽  
Canine Brucellosis

scholarly journals Rapid PCR Method for the Selection of 1,3-Pentadiene Non-Producing Debaryomyces hansenii Yeast Strains

Foods ◽  
2020 ◽  
Vol 9 (2) ◽  
pp. 162 ◽  
Author(s):  
Eva-María Rivas ◽  
Petra Wrent ◽  
María-Isabel de Silóniz
Keyword(s):  
Gene Sequence ◽  
Biocontrol Agent ◽  
Debaryomyces Hansenii ◽  
Pcr Assay ◽  
Specific Pcr ◽  
Rapid Pcr ◽  
Yeast Strains ◽  
Pcr Method ◽  
Specific Pcr Assay ◽  
Selection Of

To prevent microbial growth and its consequences, preservatives such as sorbic acid or its salts, commonly known as sorbates, are added to foods. However, some moulds and yeasts are capable of decarboxylating sorbates and producing 1,3-pentadiene. This is a volatile compound with an unpleasant “petroleum-like “odour, which causes consumer rejection of the contaminated products. In this work, we studied the production of 1,3-pentadiene in 91 strains of the yeast Debaryomyces hansenii, and we found that nearly 96% were able to produce this compound. The sequence of the FDC1Dh gene was analysed showing differences between 1,3-pentadiene producer (P) and non-producer (NP) strains. A specific PCR assay with degenerated primers based on the gene sequence was developed to discern NP and P strains. It was tested on D. hansenii strains and on some physiologically related species frequently isolated from foods, such as D. fabrii, D. subglobosus and Meyerozyma guillermondii. This method could be applied for the selection of NP D. hansenii strains, useful in biotechnological food production and as a biocontrol agent.

scholarly journals Real-Time PCR Assay for Detection and Enumeration of Dekkera bruxellensis in Wine

2003 ◽  
Vol 69 (12) ◽  
pp. 7430-7434 ◽  
Author(s):  
Trevor G. Phister ◽  
David A. Mills
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Pcr Primers ◽  
Rrna Gene ◽  
Dekkera Bruxellensis ◽  
Pcr Assay ◽  
Specific Pcr ◽  
Spoilage Yeast ◽  
Pcr Method ◽  
26S Rrna

ABSTRACT Traditional methods to detect the spoilage yeast Dekkera bruxellensis from wine involve lengthy enrichments. To overcome this difficulty, we developed a quantitative real-time PCR method to directly detect and enumerate D. bruxellensis in wine. Specific PCR primers to D. bruxellensis were designed to the 26S rRNA gene, and nontarget yeast and bacteria common to the winery environment were not amplified. The assay was linear over a range of cell concentrations (6 log units) and could detect as little as 1 cell per ml in wine. The addition of large amounts of nontarget yeasts did not impact the efficiency of the assay. This method will be helpful to identify possible routes of D. bruxellensis infection in winery environments. Moreover, the time involved in performing the assay (3 h) should enable winemakers to more quickly make wine processing decisions in order to reduce the threat of spoilage by D. bruxellensis.

Identification of Klebsiella oxytoca using a specific PCR assay targeting the polygalacturonase pehX gene

2003 ◽  
Vol 154 (8) ◽  
pp. 587-592 ◽  
Author(s):  
Gennadiy Kovtunovych ◽  
Tetyana Lytvynenko ◽  
Valentyna Negrutska ◽  
Olena Lar ◽  
Sylvain Brisse ◽  
...  
Keyword(s):  
Klebsiella Oxytoca ◽  
Pcr Assay ◽  
Specific Pcr ◽  
Specific Pcr Assay

scholarly journals Genus Level Identification of Mycobacteria from Clinical Specimens by Using an Easy-To-Handle Mycobacterium-Specific PCR Assay

1998 ◽  
Vol 36 (3) ◽  
pp. 614-617 ◽  
Author(s):  
Fritz Stauffer ◽  
Heinrich Haber ◽  
Armin Rieger ◽  
Robert Mutschlechner ◽  
Petra Hasenberger ◽  
...  
Keyword(s):  
Positive Result ◽  
Internal Control ◽  
Specific Probe ◽  
Culture Result ◽  
Pcr Assay ◽  
Clinical Specimens ◽  
Genus Level ◽  
Specific Pcr ◽  
Specific Pcr Assay ◽  
Culture Positive

An easy-to-handle Mycobacterium-specific PCR assay for detection of the presence of a wide range of mycobacterial species in clinical samples was evaluated. The performance of the genus probe was compared with the performance of probes specific forMycobacterium tuberculosis and Mycobacterium avium and with that of standard culture. In addition, the utility of an internal control in monitoring amplification inhibitors was studied. Of 545 respiratory and 325 nonrespiratory specimens (a total of 870 specimens), 58 (6.7%) showed the presence of amplification inhibitors, as determined by a negative result for the internal control. Of these 58 specimens, 31 (53%) were stool specimens; other material, even citrate blood after lysis of erythrocytes, did not pose a problem with regard to inhibition of PCR amplification. Eighty-one of the remaining 812 specimens had a positive Mycobacterium culture result. Of these culture-positive specimens, 58 (71.6%) showed a positive result with the Mycobacterium genus-specific probe. Seventy-two samples had a positive result with theMycobacterium-specific probe but a negative culture result. Of these 72 samples, 26 samples were regarded as true positive, either because the M. tuberculosis- or M. avium-specific probe was also positive at the same time or because other specimens from the same patient taken at the same time were culture positive. The sensitivity of theMycobacterium-specific probe was 78.5% and the specificity was 93.5%. This study showed that pretesting of clinical specimens for mycobacteria to the genus level with aMycobacterium-specific probe offers the routine clinical laboratory the possibility of detecting tuberculous and nontuberculous mycobacteria with one test. Furthermore, specimens testing positive with the genus-specific probe can be immediately identified with species-specific probes.

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