Detection of Riemerellosis in Ducks by gyrB Gene Based Polymerase Chain Reaction

Author(s):  
Parvathy Udayan ◽  
Priya Mani ◽  
Siniya Kollerate ◽  
Rinsha Balan ◽  
Mini Mangottumuruppel
2019 ◽  
Vol 63 (3) ◽  
pp. 27-33
Author(s):  
Š. Bursová ◽  
D. Nečasová ◽  
K. Dorotíková ◽  
L. Necidová ◽  
L. Vorlová

Abstract The aim of this study was to evaluate the toxigenic potential of Bacillus cereus strains isolated from frozen goat colostrum. Of the 50 phenotypically suspected B. cereus isolates, 39 (78.0 %) were confirmed as B. cereus by the polymerase chain reaction (PCR) method based on the gyrB gene detection. In these isolates, genes encoding the production of haemolysin BL (Hbl), a complex of non-haemolytic enterotoxins (Nhe) and emetic toxin were detected by the PCR method. In 36 (92.3 %) confirmed B. cereus isolates, genes encoding at least one type of toxins of interest were detected. In all toxigenic isolates, we found the presence of genes for Nhe production, and in 16 (41.0 %) of the isolates, genes encoding both Nhe and haemolysin BL were shown. Eight (20.5 %) of the emetic strains of B. cereus were identified. The emetic toxin production gene was always detected simultaneously with genes encoding non-haemolytic enterotoxin production. The ability to produce BL haemolysin and non-haemolytic enterotoxins were confirmed by the immunochromatographic method. In summary, goat colostrum can be a significant source of toxigenic strains of B. cereus.


Author(s):  
G. W. Hacker ◽  
I. Zehbe ◽  
J. Hainfeld ◽  
A.-H. Graf ◽  
C. Hauser-Kronberger ◽  
...  

In situ hybridization (ISH) with biotin-labeled probes is increasingly used in histology, histopathology and molecular biology, to detect genetic nucleic acid sequences of interest, such as viruses, genetic alterations and peptide-/protein-encoding messenger RNA (mRNA). In situ polymerase chain reaction (PCR) (PCR in situ hybridization = PISH) and the new in situ self-sustained sequence replication-based amplification (3SR) method even allow the detection of single copies of DNA or RNA in cytological and histological material. However, there is a number of considerable problems with the in situ PCR methods available today: False positives due to mis-priming of DNA breakdown products contained in several types of cells causing non-specific incorporation of label in direct methods, and re-diffusion artefacts of amplicons into previously negative cells have been observed. To avoid these problems, super-sensitive ISH procedures can be used, and it is well known that the sensitivity and outcome of these methods partially depend on the detection system used.


2006 ◽  
Vol 175 (4S) ◽  
pp. 485-486
Author(s):  
Sabarinath B. Nair ◽  
Christodoulos Pipinikas ◽  
Roger Kirby ◽  
Nick Carter ◽  
Christiane Fenske

1991 ◽  
Vol 66 (04) ◽  
pp. 500-504 ◽  
Author(s):  
H Peretz ◽  
U Seligsohn ◽  
E Zwang ◽  
B S Coller ◽  
P J Newman

SummarySevere Glanzmann's thrombasthenia is relatively frequent in Iraqi-Jews and Arabs residing in Israel. We have recently described the mutations responsible for the disease in Iraqi-Jews – an 11 base pair deletion in exon 12 of the glycoprotein IIIa gene, and in Arabs – a 13 base pair deletion at the AG acceptor splice site of exon 4 on the glycoprotein IIb gene. In this communication we show that the Iraqi-Jewish mutation can be identified directly by polymerase chain reaction and gel electrophoresis. With specially designed oligonucleotide primers encompassing the mutation site, an 80 base pair segment amplified in healthy controls was clearly distinguished from the 69 base pair segment produced in patients. Patients from 11 unrelated Iraqi-Jewish families had the same mutation. The Arab mutation was identified by first amplifying a DNA segment consisting of 312 base pairs in controls and of 299 base pairs in patients, and then digestion by a restriction enzyme Stu-1, which recognizes a site that is absent in the mutant gene. In controls the 312 bp segment was digested into 235 and 77 bp fragments, while in patients there was no change in the size of the amplified 299 bp segment. The mutation was found in patients from 3 out of 5 unrelated Arab families. Both Iraqi-Jewish and Arab mutations were detectable in DNA extracted from blood and urine samples. The described simple methods of identifying the mutations should be useful for detection of the numerous potential carriers among the affected kindreds and for prenatal diagnosis using DNA extracted from chorionic villi samples.


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