scholarly journals Characterization of nontypeable Actinobacillus pleuropneumoniae isolates

2020 ◽  
Vol 32 (4) ◽  
pp. 581-584
Author(s):  
Ho To ◽  
Kaho Teshima ◽  
Michiha Kon ◽  
Saori Yasuda ◽  
Yuta Akaike ◽  
...  
Keyword(s):  
Capsular Polysaccharide ◽  
Gene Clusters ◽  
Immunoblot Analysis ◽  
Actinobacillus Pleuropneumoniae ◽  
Nucleotide Sequence Analysis ◽  
Pcr Assay ◽  
Porcine Pleuropneumonia ◽  
Specific Pcr ◽  
Specific Pcr Assay

Two Actinobacillus pleuropneumoniae isolates from clinical cases of porcine pleuropneumonia in Japan were positive in the capsular serovar 15–specific PCR assay, but nontypeable (NT) in the agar gel precipitation (AGP) test. Nucleotide sequence analysis of gene clusters involved in the biosynthesis of capsular polysaccharide (CPS) and lipopolysaccharide O-polysaccharide (O-PS) revealed that both clusters contained transposable element IS Apl1 of A. pleuropneumoniae belonging to the IS30 family. Immunoblot analysis revealed that these 2 isolates could not produce O-PS. We conclude that the IS Apl1 of A. pleuropneumoniae can interfere in the biosynthesis of both CPS and O-PS.

scholarly journals Isolation and Characterization of Novel Pantoea stewartii subsp. indologenes Strains Exhibiting Center Rot in Onion

Plant Disease ◽  
2018 ◽  
Vol 102 (4) ◽  
pp. 727-733 ◽  
Author(s):  
Spencer Stumpf ◽  
Brian Kvitko ◽  
Ron Gitaitis ◽  
Bhabesh Dutta
Keyword(s):  
Pearl Millet ◽  
Phylogenetic Analyses ◽  
Bacterial Strains ◽  
Pcr Assay ◽  
Pantoea Stewartii ◽  
Specific Pcr ◽  
Isolation And Characterization ◽  
Distinct Cluster ◽  
Specific Pcr Assay

Center rot of onion is an economically important disease caused by three Pantoea spp.: Pantoea ananatis, P. agglomerans, and P. allii. Symptoms caused by these three species are similar and include white streaking and necrosis of foliage; and, in some cases, the bacterium may enter the bulb, causing liquefaction and rot of bulb scales. Two bacterial strains were isolated from onion expressing symptoms indicative of center rot from two different outbreaks in Toombs County, GA in 2003 (PNA 03-3) and 2014 (PNA 14-12). These strains were initially identified as P. ananatis based on physiological and specific polymerase chain reaction (PCR) assays; however, further 16S ribosomal RNA (rRNA) and multilocus sequence analysis showed that the strains were more closely related to P. stewartii subsp. stewartii and P. stewartii subsp. indologenes. Further characterization using phylogenetic analysis, a P. stewartii subsp. indologenes-specific PCR assay, indole test, and pathogenicity on onion and pearl millet were conducted. Phylogenetic analyses (16S rRNA and atpD, gyB, infB, and rpoB genes) revealed that these strains formed a distinct cluster with the type strains of P. stewartii subsp. indologenes LMG 2632T and P. stewartii subsp. stewartii LMG 2715T separate from P. ananatis, P. agglomerans, and P. allii. Furthermore, onion strains were amplified with the P. stewartii subsp. indologenes-specific PCR assay. The pathogenicity assays with onion strains showed that they were pathogenic on onion and pearl millet, a known host of P. stewartii subsp. indologenes. However, the type strain of P. stewartii subsp. indologenes LMG 2632T was pathogenic only on pearl millet but not on onion. These results suggest that the onion strains PNA 03-3 and PNA 14-12 can potentially be novel P. stewartii subsp. indologenes strains capable of producing symptoms on onion. Hence, we recommend the inclusion of P. stewartii subsp. indologenes as the fourth member in the center rot complex of onion, along with P. ananatis, P. agglomerans, and P. allii.

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.

scholarly journals Evaluation of a vanA-Specific PCR Assay for Detection of Vancomycin-Resistant Enterococcus faecium during a Hospital Outbreak

1999 ◽  
Vol 37 (10) ◽  
pp. 3348-3349 ◽  
Author(s):  
Michel Roger ◽  
Marie-Claude Faucher ◽  
Pierre Forest ◽  
Pierre St-Antoine ◽  
François Coutlée
Keyword(s):  
Enterococcus Faecium ◽  
Pcr Assay ◽  
Culture Methods ◽  
Agar Culture ◽  
Enrichment Broth ◽  
Specific Pcr ◽  
Hospital Outbreak ◽  
Vancomycin Resistant Enterococci ◽  
Vancomycin Resistant ◽  
Specific Pcr Assay

We investigated the use of PCR as an alternative to culture of fecal samples for detection of vanA-containingEnterococcus faecium during a recent hospital outbreak. Rectal swabs collected consecutively from 223 patients were analyzed by culture with and without enrichment broth and byvanA-specific PCR of enrichment broth samples. Fifty-five specimens were positive for vanA-containing E. faecium by at least one method. The sensitivities of thevanA-specific PCR assay and agar culture with and without enrichment broth were 94.5, 98, and 89%, respectively. All three methods were 100% specific. Final results were obtained much more rapidly by PCR (within 24 to 30 h of specimen submission) than by the culture methods (4 to 5 days). Thus, PCR is an accurate and rapid alternative to culture for detection of vancomycin-resistant enterococci during hospital outbreaks.

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

Acta Tropica ◽  
2006 ◽  
Vol 100 (3) ◽  
pp. 241-245 ◽  
Author(s):  
Mallorie Hide ◽  
Anne-Laure Bañuls
Keyword(s):  
Pcr Assay ◽  
Specific Pcr ◽  
Specific Pcr Assay ◽  
Species Specific
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