scholarly journals Biotyping and Antimicrobial Susceptibility of Enterococcus faecalis and E. faecium Isolated from Urine and Stool Samples

2021 ◽  
Vol 13 (10) ◽  
Author(s):  
Gülşah Tollu ◽  
Ismail Hakkı Ekin
Keyword(s):  
Multiplex Pcr ◽  
Antimicrobial Susceptibility ◽  
Animal Health ◽  
Molecular Techniques ◽  
Penicillin G ◽  
Clinical Approach ◽  
Biochemical Tests ◽  
Specific Primers ◽  
Stool Samples ◽  
Species Specific

Background: Enterococci are one of the opportunistic pathogenic microorganisms that can cause significant problems for human and animal health. Enterococcus faecium seems to be more resistant to antibiotics than E. faecalis. It is thought that pathogenic E. faecium can develop antibiotic resistance very quickly, and the ability to transfer this feature is considered to be an important health risk. Objectives: This study aimed to determine the prevalence, biotypes, and in vitro antimicrobial susceptibility of E. faecalis and E. faecium strains isolated from 267 routine urine and stool samples that were brought to the microbiology laboratory of Regional Training and Research Hospital of Van, with permission of the patients. Methods: In the present study, enterococci using species-specific primers to examine E. faecalis and E. faecium multiplex PCR technique was applied. Biotyping of the isolates was used to identify them as E. faecalis and E. faecium by molecular techniques, and antibiotic susceptibility of all samples was examined, as well. Results: The isolates were identified by multiplex PCR using species-specific primers for E. faecalis and E. faecium. Biotyping based on 13 biochemical tests showed that 72.5%, 12.5%, and 15% of E. faecalis strains were of biotypes I, II, and III, respectively, whereas E. faecium strains could be divided into biotype I (10%), biotype II (12.5%), biotype III (27.5%), and biotype IV (50%). Additionally, all E. faecalis strains were found to be susceptible to penicillin G and imipenem. On the other hand, 95% of the E. faecalis strains were found to be resistant to clindamycin, 77.5% to tetracycline and trimethoprim/sulfamethoxazole, 42.5% to erythromycin, 32.5% to gentamicin, and 17.5% to ciprofloxacin. Of E. faecium strains, 37.5% were found to be resistant to clindamycin, 32.5% to penicillin G, 27.5% to erythromycin and imipenem, 20% to ciprofloxacin, 17.5% to tetracycline and trimethoprim/sulfamethoxazole, 15% to gentamicin, and 5% to vancomycin. Conclusions: In conclusion, the identification of E. faecalis and E. faecium strains by PCR is reliable and faster than biochemical tests. Additionally, the results of antimicrobial susceptibility tests may provide important contributions to the clinical approach.

Development of a multiplex PCR assay for the detection of key genes associated with Pasteurella multocida subspecies

2021 ◽  
pp. 104063872110634
Author(s):  
Barbara Ujvári ◽  
Hubert Gantelet ◽  
Tibor Magyar
Keyword(s):  
Multiplex Pcr ◽  
Pasteurella Multocida ◽  
Rrna Gene ◽  
Pcr Assay ◽  
Biochemical Tests ◽  
Specific Primers ◽  
Multiplex Pcr Assay ◽  
Subspecies Level ◽  
Species Specific ◽  
Reference Strains

The ability to distinguish among the subspecies of Pasteurella multocida isolates is important epidemiologically; however, classification at the subspecies level based on the results of conventional biochemical tests (fermentation of sorbitol and dulcitol) is reportedly not accurate in all cases. Therefore, we developed a rapid, multiplex PCR assay to differentiate among the 3 subspecies of P. multocida. The PCR assay includes the P. multocida species–specific primers KMT1SP6 and KMT1T7 as an internal amplification control, with a newly designed gatD (galactitol-1-phosphate-5-dehydrogenase)-specific primer pair (unique for subsp. gallicida), and primers targeting a 16S rRNA gene region specific for subsp. septica. The subspecies specificity of the PCR was demonstrated by applying the test to a collection of 70 P. multocida isolates, including the Heddleston serovar reference strains; all isolates and strains were assigned correctly. The PCR assay is a sensitive, specific, and highly effective method for the identification of P. multocida subspecies, and an alternative to biochemical test–based differentiation. A possible relationship was noticed between P. multocida subspecies and lipopolysaccharide (LPS) genotype; all but one of the subsp. gallicida strains were isolated only from avian hosts and represented L1 LPS genotype. Subsp. multocida and subsp. septica isolates were classified into 5 and 4 different LPS genotypes, respectively, of which L3 was the only LPS genotype shared between these 2 subspecies.

scholarly journals Antimicrobial Resistance of Enterococcus Faecium Isolated from the Urinary System Of Dogs

2019 ◽  
Vol 42 (1) ◽  
pp. 15-21 ◽  
Author(s):  
Sukru Kirkan ◽  
Ugur Parin ◽  
Gamze Balat
Keyword(s):  
Antimicrobial Resistance ◽  
Multiplex Pcr ◽  
Antimicrobial Susceptibility ◽  
Enterococcus Faecium ◽  
Vancomycin Resistance ◽  
Urine Samples ◽  
Urinary System ◽  
Specific Primers ◽  
Vancomycin Resistant ◽  
Species Specific

AbstractThe purpose of the present study was to determine the antimicrobial susceptibility of vancomycin-resistant and Enterococcus faecium strains isolated from urine samples of dogs. A total of 22 Enterococcus sp. samples were isolated and identified from 100 urine samples collected by cystocentesis from dogs of both sexes. The identification with species specific primers for multiplex PCR revealed that all 22 isolates (100%) belonged to E. faecium. Vancomycin resistance was found in 10 (45%) samples of E. faecium strains with PCR study by vanA and vanB primers.

scholarly journals Multiplex PCR with 16S rRNA Gene-Targeted Primers of Bifidobacterium spp. To Identify Sources of Fecal Pollution

2004 ◽  
Vol 70 (5) ◽  
pp. 3171-3175 ◽  
Author(s):  
X. Bonjoch ◽  
E. Ballesté ◽  
A. R. Blanch
Keyword(s):  
16S Rrna ◽  
Multiplex Pcr ◽  
Fecal Pollution ◽  
Sensitivity Threshold ◽  
Rrna Gene ◽  
Specific Primers ◽  
Animal Wastewater ◽  
Wastewater Samples ◽  
Species Specific ◽  
Different Origins

ABSTRACT Bifidobacteria are one of the most common bacterial types found in the intestines of humans and other animals and may be used as indicators of human fecal pollution. The presence of nine human-related Bifidobacterium species was analyzed in human and animal wastewater samples of different origins by using species-specific primers based on 16S rRNA sequences. Only B. adolescentis and B. dentium were found exclusively in human sewage. A multiplex PCR approach with strain-specific primers was developed. The method showed a sensitivity threshold of 10 cells/ml. This new molecular method could provide useful information for the characterization of fecal pollution sources.

Microsporidial infections due toEncephalitozoon intestinalisin non-HIV-infected patients with chronic diarrhoea

2011 ◽  
Vol 140 (10) ◽  
pp. 1773-1779 ◽  
Author(s):  
J. YAKOOB ◽  
Z. ABBAS ◽  
M. ASIM BEG ◽  
W. JAFRI ◽  
S. NAZ ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Stool Examination ◽  
Chronic Diarrhoea ◽  
Healthy Controls ◽  
Chain Reaction ◽  
Specific Primers ◽  
Specific Pcr ◽  
Stool Samples ◽  
Polymerase Chain ◽  
Species Specific

SUMMARYWe determined the prevalence of microsporidiaEnterocytozoon(Ent.)bieneusiandEncephalitozoon(E.)intestinalisinfection in patients with chronic diarrhoea and hepatocellular carcinoma (HCC). A total of 330 stool samples were examined from 171 (52%) patients with chronic diarrhoea, 18 (5%) with HCC while 141 (43%) were controls. Stool microscopy, polymerase chain reaction (PCR) with species-specific primers forEnt. bieneusiandE. intestinalisand sequencing were carried out. Microsporidia were found by trichrome staining in 11/330 (3%) andE. intestinalisby PCR in 13/330 (4%) whileEnt. bieneusiwas not detected. PCR forE. intestinaliswas positive in 8/171 (5%) stool samples from patients with chronic diarrhoea, 2/141 (1·4%) samples from healthy controls and in 3/18 (17%) samples from patients with HCC. In the chronic diarrhoea group,E. intestinaliswas positive in 4/171 (2·3%) (P=0·69) stool samples compared to 2/18 (11%) (P=0·06) in the HCC group and 2/141 (1·4%) from healthy controls.E. intestinalisinfection was significantly associated with chronic diarrhoea and HCC in these patients who were negative for HIV. Stool examination with trichrome or species-specific PCR for microsporidia may help establish the cause of chronic diarrhoea.

scholarly journals Detection of Leishmania (L.) infantum in stray dogs by molecular techniques with sensitive species-specific primers

2016 ◽  
Vol 37 (1) ◽  
pp. 23-30 ◽  
Author(s):  
Rodrigo C. Silva ◽  
Virgínia B. Richini-Pereira ◽  
Mariana Kikuti ◽  
Pâmela M. Marson ◽  
Helio Langoni
Keyword(s):  
Molecular Techniques ◽  
Sensitive Species ◽  
Specific Primers ◽  
Stray Dogs ◽  
Species Specific

scholarly journals Diversity of Colletotrichum Species causing Anthracnose Disease from Mango cv. Nam Dork Mai See Tong based on ISSR-PCR

2021 ◽  
Author(s):  
S. Bincader ◽  
R. Pongpisutta ◽  
C. Rattanakreetakul
Keyword(s):  
Species Complex ◽  
Pcr Amplification ◽  
Molecular Techniques ◽  
Specific Primers ◽  
Cophenetic Correlation ◽  
Anthracnose Disease ◽  
Colletotrichum Species ◽  
Morphological Studies ◽  
Pathogenicity Tests ◽  
Species Specific

Background: Anthracnose disease caused by the genus Colletotrichum is one of the crucial problems occurring in the field, along with postharvest diseases and affects mango quality in Thailand. In particular, the Nam Dork Mai See Tong cultivar, which is highly susceptible to the disease, is an important product for exportation. Methods: In this research, thirty-seven Colletotrichum species isolate were obtained from anthracnose disease in mango cv. Nam Dork Mai See Tong in three provinces in Thailand. Morphological studies and molecular techniques using species-specific primers were investigated; moreover, the diversity of pathogens was analyzed using PCR amplification of inter simple sequence repeats (ISSRs) with 6 primers, including pathogenicity tests. Result: Morphological studies and molecular detection with species-specific primers revealed that 32 isolates belonged to the C. gloeosporioides species complex and 5 isolates to the C. acutatum species complex. The genetic diversity of pathogens was analyzed. PCR amplification using 6 ISSR primers produced 35 polymorphic bands. These bands were used to construct UPGMA, in which cluster analysis divided the 37 isolates into 3 main groups and 8 subgroups at 61-73% Jaccard similarity coefficient with cophenetic correlation (r) = 0.6781. The ISSR technique showed the greatest genetic variation among isolates collected from different locations. Hence, a study based on ISSR markers was profitable to investigate the phylogenetic relationship of the genus Colletotrichum. Pathogenicity tests revealed that PC006 (Ca) and CS005 (Cg) showed the highest aggressiveness, with disease incidences of 84.74 and 80.90%, respectively. This study indicates that the diversity of pathogenic Colletotrichum species related to mango plantations in Thailand is increasing.

Agronomically important thrips: development of species-specific primers in multiplex PCR and microarray assay using internal transcribed spacer 1 (ITS1) sequences for identification

2014 ◽  
Vol 105 (1) ◽  
pp. 52-59 ◽  
Author(s):  
W.B. Yeh ◽  
M.J. Tseng ◽  
N.T. Chang ◽  
S.Y. Wu ◽  
Y.S. Tsai
Keyword(s):  
Multiplex Pcr ◽  
Species Identification ◽  
Internal Transcribed Spacer ◽  
High Throughput Screening ◽  
Virus Transmission ◽  
Specific Primers ◽  
Its1 Sequence ◽  
Microarray Assay ◽  
Thrips Species ◽  
Species Specific

AbstractThrips, the sole vector of plantTospovirus, are major pests of many agricultural crops throughout the world. Molecular approaches have been applied in recent decades to identify these minute and morphologically difficult to distinguish insects. In this study, sequences of internal transcribed spacer 1 (ITS1) region of 15 agronomically important thrips, including several virus transmission species, have been analyzed in order to design species-specific primers for multiplex PCR and probes for microarray assay. That the ITS1 sequence distances within species were smaller than those among species suggests that the ITS1 fragment can be used for thrips species identification. The specificity and stability of these primers, combined with universal paired primers, were tested and verified in multiplex PCR. Using these specific primers as probes, microarray assay showed that PCR products of all thrips species hybridized consistently to their corresponding probes, though some signals were weak. We have demonstrated that multiplex PCR using specific primers based on ITS1 sequences is a simple, reliable, and cost-effective diagnostic tool for thrips species identification. Moreover, the DNA microarray assay is expected to extend into a reliable high-throughput screening tool for the vast numbers of thrips.

scholarly journals Molecular identification of Atlantic goliath grouper Epinephelus itajara (Lichtenstein, 1822) (Perciformes: Epinephelidae) and related commercial species applying multiplex PCR

2016 ◽  
Vol 14 (3) ◽  
Author(s):  
Júnio S. Damasceno ◽  
Raquel Siccha-Ramirez ◽  
Claudio Oliveira ◽  
Fernando F. Mendonça ◽  
Arthur C. Lima ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Cost Effective ◽  
Molecular Techniques ◽  
Forensic Identification ◽  
Correct Identification ◽  
Western Atlantic ◽  
Endangered Fish ◽  
Critically Endangered Species ◽  
Goliath Grouper ◽  
Species Specific

ABSTRACT The Atlantic goliath grouper, Epinephelus itajara , is a critically endangered species, threatened by illegal fishing and the destruction of its habitats. A number of other closely related grouper species found in the western Atlantic are also fished intensively. While some countries apply rigorous legislation, illegal harvesting followed by the falsification of fish products, which impedes the correct identification of the species, is a common practice, allowing the catch to be marketed as a different grouper species. In this case, molecular techniques represent an important tool for the monitoring and regulation of fishery practices, and are essential for the forensic identification of a number of different species. In the present study, species-specific primers were developed for the Cytochrome Oxidase subunit I gene, which were applied in a multiplex PCR for the simultaneous identification of nine different species of Epinephelidae: Epinephelus itajara , E. quinquefasciatus , E. morio , Hyporthodus flavolimbatus , H. niveatus , Mycteroperca acutirostris , M. bonaci , M. marginata , and M. microlepis . Multiplex PCR is a rapid, reliable and cost-effective procedure for the identification of commercially-valuable endangered fish species, and may represent a valuable tool for the regulation and sustainable management of fishery resources.

Sign in / Sign up

Export Citation Format

Share Document