MOLECULAR TYPING OF METHICILLIN-SUSCEPTIBLE STAPHYLOCOCCUS AUREUS (MSSA), ISOLATED FROM MONKEYS, BASED ON COAGULASE GENE POLYMORPHISM

2019 ◽  
pp. 57-62
Author(s):  
V. A. Kalashnikova
Keyword(s):  
Staphylococcus Aureus ◽  
Real Time ◽  
Real Time Pcr ◽  
Molecular Genetic ◽  
Bovine Mastitis ◽  
Variable Region ◽  
Staphylococcal Infection ◽  
Pcr Analysis ◽  
Polymorphism Analysis ◽  
Coa Gene

Staphylococcus aureus is a very dangerous microorganism that causes more than 100 nosological forms of disease in humans and animals, including pneumonia, skin and soft tissue infections, food toxicoinfections, wound abscess, etc. Numerous studies on genotyping Staphylococcus aureus, isolated from humans, food and bovine mastitis have been carried out. The lack of information on the genotyping of these pathogens detected in monkeys living in captivity served as a stimulus to conduct a similar research, since staphylococcal infections in the primates are widespread. The present study is devoted to the study of the polymorphism of a variable region of the coagulase gene and to the typing of Staphylococcus aureus isolates from monkeys of different species kept at Adler monkey farm. 115 Staphylococcus aureus isolates were studied using phenotypic and molecular genetic methods. Genotyping was performed using PCR, real-time PCR and PCR with subsequent restriction fragment length polymorphism analysis (PCR-RFLP). A real-time PCR analysis allowed to classify all Staphylococcus aureus as methicillin-susceptible staphylococci (MSSA). After amplification of a variable region of the coagulase gene, 4 types of amplicons of 600, 700, 800, and 900 bp were generated. This data demonstrates structural differences of this gene in the studied isolates. The coagulase gene of 900 bp prevailed. The use of the Cfo1 endonuclease allowed to identify 23 different restriction profiles of the coa gene, but only three of them predominated. Staphylococcus aureus bacteria with seven types of coagulase gene were found only in the lungs of monkeys that died of pneumonia. The results obtained suggest that these isolates have tropism for lung tissue. Among Staphylococcus aureus isolated from pneumonia cases, isolates with three types of the coa gene prevailed. Staphylococcus aureus of eleven types of coagulase gene can be attributed to the invasive isolates, since they were detected in the tissues of various organs. Staphylococcal infection in monkeys kept at the monkey farm is caused by genotypically heterogeneous population of Staphylococcus aureus.

Gene-Related Strain Variation of Staphylococcus aureus for Homologous Resistance Response to Acid Stress

2014 ◽  
Vol 77 (10) ◽  
pp. 1794-1798 ◽  
Author(s):  
SOOMIN LEE ◽  
SOOYEON AHN ◽  
HEEYOUNG LEE ◽  
WON-IL KIM ◽  
HWANG-YONG KIM ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Real Time ◽  
Real Time Pcr ◽  
Acid Resistance ◽  
Transcriptional Analysis ◽  
Pcr Analysis ◽  
Quantitative Real Time Pcr ◽  
Resistance Response ◽  
Expression Levels ◽  
Acid Shock

This study investigated the effect of adaptation of Staphylococcus aureus strains to the acidic condition of tomato in response to environmental stresses, such as heat and acid. S. aureus ATCC 13565, ATCC 14458, ATCC 23235, ATCC 27664, and NCCP10826 habituated in tomato extract at 35°C for 24 h were inoculated in tryptic soy broth. The culture suspensions were then subjected to heat challenge or acid challenge at 60°C and pH 3.0, respectively, for 60 min. In addition, transcriptional analysis using quantitative real-time PCR was performed to evaluate the expression level of acid-shock genes, such as clpB, zwf, nuoF, and gnd, from five S. aureus strains after the acid habituation of strains in tomato at 35°C for 15 min and 60 min in comparison with that of the nonhabituated strains. In comparison with the nonhabituated strains, the five tomato-habituated S. aureus strains did not show cross protection to heat, but tomato-habituated S. aureus ATCC 23235 showed acid resistance. In quantitative real-time-PCR analysis, the relative expression levels of acid-shock genes (clpB, zwf, nuoF, and gnd) were increased the most in S. aureus ATCC 23235 after 60 min of tomato habituation, but there was little difference in the expression levels among the five S. aureus strains after 15 min of tomato habituation. These results indicate that the variation of acid resistance of S. aureus is related to the expression of acid-shock genes during acid habituation.

scholarly journals Regulation of virulence and β-lactamase gene expression in Staphylococcus aureus isolates: cooperation of two-component systems in bloodstream superbugs

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Sanaz Dehbashi ◽  
Hamed Tahmasebi ◽  
Behrouz Zeyni ◽  
Mohammad Reza Arabestani
Keyword(s):  
Staphylococcus Aureus ◽  
Real Time ◽  
Real Time Pcr ◽  
Quantitative Real Time Pcr ◽  
Gene Expressions ◽  
Resistance Development ◽  
Component Systems ◽  
Two Component ◽  
Two Component Systems ◽  
Lactamase Gene

Abstract Background Methicillin-resistant Staphylococcus aureus (MRSA)-bloodstream infections (BSI) are predominantly seen in the hospital or healthcare-associated host. Nevertheless, the interactions of virulence factor (VFs) regulators and β-lactam resistance in MRSA-BSI are unclear. This study aims to characterize the molecular relationship of two-component systems of VFs and the expression of the β-lactamase gene in MRSA-BSI isolates. In this study, 639 samples were collected from BSI and identified by phenotypic methods. We performed extensive molecular characterization, including SCCmec type, agr type, VFs gene profiles determinations, and MLST on isolates. Also, a quantitative real-time PCR (q-RT PCR) assay was developed for identifying the gene expressions. Results Ninety-one (91) S. aureus and 61 MRSA (67.0%) strains were detected in BSI samples. The presence of VFs and SCCmec genes in MRSA isolates were as follows: tst (31.4%), etA (18.0%), etB (8.19%), lukS-PVL (31.4%), lukF-PV (18.0%), lukE-lukD (16.3%), edin (3.2%), hla (16.3%), hlb (18.0%), hld (14.7%), hlg (22.9%), SCCmecI (16.3%), SCCmecII (22.9%), SCCmecIII (36.0%), SCCmecIV (21.3%), and SCCmecV (16.3%). Quantitative real-time PCR showed overexpression of mecRI and mecI in the toxigenic isolates. Moreover, RNAIII and sarA genes were the highest expressions of MRSA strains. The multi-locus sequence typing data confirmed a high prevalence of CC5, CC8, and CC30. However, ST30, ST22, and ST5 were the most prevalent in the resistant and toxigenic strains. Conclusion We demonstrated that although regulation of β-lactamase gene expressions is a significant contributor to resistance development, two-component systems also influence antibiotic resistance development in MRSA-BSI isolates. This indicates that resistant strains might have pathogenic potential. We also confirmed that some MLST types are more successful colonizers with a potential for MRSA-BSI.

ID: 316 Real-time PCR analysis of BCL2L12, a novel member of BCL2 family of the apoptosis-related genes, in human leukemia.

2006 ◽  
Vol 4 (s1) ◽  
pp. 82-82
Author(s):  
K. Floros ◽  
H. Thomadaki ◽  
S. Pavlovic ◽  
M. Talieri ◽  
M. Colovic ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Pcr Analysis ◽  
Human Leukemia ◽  
Bcl2 Family

Improved diagnosis of gastrointestinal infections using a semi-automated multiplex real-time PCR for detection of enteropathogens

2021 ◽  
Vol 70 (9) ◽  
Author(s):  
Berta Fidalgo ◽  
Elisa Rubio ◽  
Victor Pastor ◽  
Marta Parera ◽  
Clara Ballesté-Delpierre ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Enteric Pathogens ◽  
Pcr Analysis ◽  
Culture Methods ◽  
Gastrointestinal Infections ◽  
Content Type ◽  
Link Type ◽  
Microbiological Culture ◽  
Micro Organisms

Introduction. The identification of enteropathogens is critical for the clinical management of patients with suspected gastrointestinal infection. The FLOW multiplex PCR system (FMPS) is a semi-automated platform (FLOW System, Roche) for multiplex real-time PCR analysis. Hypothesis/Gap Statement. FMPS has greater sensitivity for the detection of enteric pathogens than standard methods such as culture, biochemical identification, immunochromatography or microscopic examination. Aim.The diagnostic performance of the FMPS was evaluated and compared to that of traditional microbiological procedures. Methodology. A total of 10 659 samples were collected and analysed over a period of 7 years. From 2013 to 2018 (every July to September), samples were processed using standard microbiological culture methods. In 2019, the FMPS was implemented using real-time PCR to detect the following enteropathogens: Shigella spp., Salmonella spp., Campylobacter spp., Giardia intestinalis, Entamoeba histolytica, Blastocystis hominis, Cryptosporidum spp., Dientamoeba fragilis, adenovirus, norovirus and rotavirus. Standard microbiological culture methods (2013–2018) included stool culture, microscopy and immunochromatography. Results. A total of 1078 stool samples were analysed prospectively using the FMPS from July to September (2019): bacterial, parasitic and viral pathogens were identified in 15.3, 9.71 and 5.29 % of cases, respectively. During the same period of 6 years (2013–2018), the proportion of positive identifications using standard microbiological methods from 2013 to 2018 was significantly lower. A major significant recovery improvement was observed for all bacteria species tested: Shigella spp./enteroinvasive Escherichia coli (EIEC) (P <0.05), Salmonella spp. (P <0.05) and Campylobacter spp. (P <0.05). Marked differences were also observed for the parasites G. intestinalis, Cryptosporidium spp. and D. fragilis. Conclusion. These results support the value of multiplex real-time PCR analysis for the detection of enteric pathogens in laboratory diagnosis with outstanding performance in identifying labile micro-organisms. The identification of unsuspected micro-organisms for less specific clinical presentations may also impact on clinical practice and help optimize patient management.

scholarly journals Real-time PCR analysis of the apoptosis related genes in ATRA treated APL t(15;17) patients

2003 ◽  
Vol 35 (5) ◽  
pp. 454-459 ◽  
Author(s):  
Hakan Savli ◽  
Sema Sirma ◽  
Balint Nagy ◽  
Melih Aktan ◽  
Guncag Dincol ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Pcr Analysis
Sign in / Sign up

Export Citation Format

Share Document