scholarly journals Application of Multiplex PCR for the Identification of Oxacillinase Genes and Determination of Antibiotic Resistance Pattern in Environmental Isolates of Acinetobacter baumannii in ICU

2021 ◽  
Vol 8 (3) ◽  
pp. 89-93
Author(s):  
Anahita Farajzadeh ◽  
Mohsen Mirzaee ◽  
Shahram Nanekarani ◽  
Reza Yari
Keyword(s):  
Acinetobacter Baumannii ◽  
Multiplex Pcr ◽  
Polymyxin B ◽  
Resistance Pattern ◽  
Rrna Gene ◽  
Environmental Isolates ◽  
Biochemical Tests ◽  
Cross Sectional ◽  
Significance Level ◽  
Genes Encoding

Background: Acinetobacter baumannii is a common cause of nosocomial infections. A prominent feature of these bacteria is resistance to carbapenems. This study aimed to identify OXA genes encoding oxacillinase in Acinetobacter baumannii isolates. Methods: This cross-sectional descriptive study was performed on 25 environmental A. baumannii isolates collected from ICU over 8 months. Definitive identification of isolates was performed by biochemical tests and polymerase chain reaction (PCR) of 16s rRNA gene. Antibiotic susceptibility testing was performed on Müller-Hinton agar medium by disk diffusion and E-test. Antibiogram and multiplex PCR data of beta-lactamase genes were collected and analyzed at a significance level of P<0.05 using SPSS 22.0. Results: Except for one isolate, all isolates (96%) were sensitive to polymyxin B and 80% of isolates were sensitive to oxacillin. All isolates were sensitive to meropenem, ampicillin/sulbactam, gentamicin, amikacin, piperacillin, and carbenicillin. The results showed that 25 isolates (100%) had OXA-51 gene, 21 isolates (84%) had OXA-58 gene, one isolate (4%) had OXA-24 gene, and none of the isolates contained OXA-23 gene. Only isolate No.10 had three oxacillinase genes simultaneously and it was resistant to oxacillin, polymyxin B, and cephalothin. Conclusions: The study showed that environmental isolates of ICU do not have pathogenic genes present in the clinical isolates, and how these genes are transferred to the peripheral isolates is an important point that should be studied. Identification of genes encoding carbapenem resistance may help to understand the mechanisms of resistance transfer in A. baumannii. The lack of the OXA-23 gene plays an important role in the susceptibility of isolates to antibiotics and non-emergence of resistant strains.

scholarly journals Identification through Culture and Molecular Methods of Campylobacter jejuni, Campylobacter coli and Campylobacter fetus in Surface Waters in Rasht

2019 ◽  
pp. 1-7
Author(s):  
Keyvan Roshanjo ◽  
Nematallah Jonaidi Jafari ◽  
Leila Asadpour ◽  
Reza Ranjbar ◽  
Davoud Afshar ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Campylobacter Jejuni ◽  
Surface Waters ◽  
Disease Transmission ◽  
Cross Sectional Study ◽  
Campylobacter Coli ◽  
Infectious Agents ◽  
Biochemical Tests ◽  
Cross Sectional ◽  
Campylobacter Fetus

Backgrounds: As zoonotic infectious agents, Campylobacter spp. are important factors causing gastroenteritis in humans. Surveys show that the three strains; Campylobacter jejuni, Campylobacter coli and Campylobacter fetus play a major role in human infections. Identification of these infectious agents is valuable for sanitary control of disease transmission through water resources. Objectives: The aim of this study was identification and molecular diagnosis of Campylobacter jejuni, Campylobacter coli and Campylobacter fetus in surface waters in Rasht. Materials and Methods: This cross-sectional study was conducted on 45 samples of surface water in Rasht collected according to water health guidelines. After culture and biochemical tests on collected samples, detection and identification of Campylobacter jejuni, Campylobacter coli and Campylobacter fetus was done using sequence-specific amplification by Multiplex PCR. The results were subjected to statistical analysis using SPSS software. Results: Out of 45 samples tested, 6 were positive in culture, four of which were identified as Campylobacter jejuni after biochemical tests. Using Multiplex PCR, 8 samples were positive, from which 3 were Campylobacter jejuni, 1 Campylobacter coli and 4 were positive for both Campylobacter jejuni and Campylobacter coli. All the samples did not yield C. fetus. Conclusions: Multiplex PCR is regarded a diagnostic method with higher sensitivity and specificity than compared to methods for Campylobacter. The prevalence of Campylobacter jejuni and Campylobacter coli in surface waters in Rasht is considerable. Therefore, public health measures for the control of these organisms are recommended.

scholarly journals Characteristic and Potential Therapeutic Effect of Isolated MDR-Acinetobacter Baumannii Lytic Phage

2020 ◽  
Author(s):  
Behnam Sisakhtpour ◽  
Arezoo Mirzaei ◽  
Vajihe Karbasizadeh ◽  
Nafise Sadat Hosseini ◽  
Mehdi Shabani ◽  
...  
Keyword(s):  
Acinetobacter Baumannii ◽  
Bacterial Resistance ◽  
Cross Sectional Study ◽  
Resistance Pattern ◽  
Lytic Phage ◽  
Klebsiella Pneumonia ◽  
Cross Sectional ◽  
Coomassie Blue ◽  
Phage Sensitivity ◽  
Multiple Drugs

Abstract Background: Acinetobacter baumannii is a major pathogen in the hospital, especially in Intensive Care Units (ICU) and the resistance to multiple drugs as a major contributor to hospital infection. Bacteriophages are viruses that attack bacteria and kill them that could be used for clinical treatment. The aim of the study is in evaluating the function of bacteriophage specificity of multi-drug resistant Acinetobacter baumannii, to be used as a useful method for treating of Acinetobacter Infections.Methods: Cross-sectional study during the year 2017, from patients admitted to the ICU, First, 48 isolates of Acinetobacter baumannii were identified by phenotypic method and amplified with blaOXA-51 gene. Then, the sensitivity of phages to pathogens namely ESKAPE (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumonia, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter spp) evaluated. DNA of the phage was extracted using the Viral Nucleic Extraction Kit II (Geneaid, Taipei, Taiwan) according to the manufacturer's instructions. Then for protein analysis, PEG-precipitated purified phages were subjected directly to SDS-PAGE, and protein bands were visualized by coomassie Blue G-250 staining method. Finally for cell survival assay we investigated the toxicity of the isolated phage to Hela cells.Results: In the bacterial resistance pattern, the highest resistance belongs to ciprofloxacin. In optimal phage test, at dilution of 1 (MOI 1) it produced the best effect on bacteria in 30 minutes. Phage sensitivity to different hosts performed by double layer agar method, the phage was treated with ESKAPE bacteria and after 24 hours’ incubation at 37°C, only for Acinetobacter baumannii Plaque created. The genome analysis indicated that phage pIsf-AB2 has a double-stranded DNA genome. In bacterial control, all cells were killed by A. baumannii, and no live-cell was seen. The cells remained in control of the phage, and the phage did not affect the cells.Conclusion: Our findings support the potential application of the phage with potent endolysin activity against MDR A. baumannii and give useful information for its further study and use.

scholarly journals Prevalence and distribuion of staphylococcal enterotoxin genes among Staphylococcus aureus isolates from chicken and turkey carcasses in Algeria

2019 ◽  
Vol 69 (4) ◽  
pp. 1297 ◽  
Author(s):  
F. Mebkhout ◽  
L. Mezali ◽  
T. M. Hamdi ◽  
Z. Cantekin ◽  
Y. Ergun ◽  
...  
Keyword(s):  
Public Health ◽  
Staphylococcus Aureus ◽  
Multiplex Pcr ◽  
Staphylococcal Enterotoxin ◽  
Foodborne Illnesses ◽  
Biochemical Tests ◽  
Staphylococcal Enterotoxins ◽  
Enterotoxin Genes ◽  
Real Risk ◽  
Genes Encoding

This study is aimed to determine the prevalence of staphylococcus aureus (S.aureus) by biochemical tests in poultry carcasses. It is also intend to detect the distribution of genes for classical staphylococcal enterotoxins A, B, C, D and E (sea, seb, sec, sed and see) and for gene femA, specific for S.aureus species, using multiplex PCR. A total of 385 samples of neck skins from fresh poultry carcasses were collected during the period 2012-2013 from 16 different slaughterhouses located in the region of Algiers, Algeria. The overall prevalence of S.aureus in freshly slaughtered poultry carcasses was 41.56%, with an individual prevalence of 40.63% and 45.71% for chicken and turkey respectively. From the 95 strains of S.aureus identified by biochemical tests, 82 (86.32%) isolates were femA positive using multiplex PCR. The investigation has also revealed the presence of both enterotoxins B and D, with a predominance of seb (13.33%) followed by sed (1.67%), in the chicken carcasses while in turkey only sed was detected (4.55%) It has been found that strains of S.aureus of poultry origin can be enterotoxigenic with the predominance of genes encoding for enterotoxins seb in chicken and sed in turkey. As enterotoxins can be produced in adequate amounts to induce foodborne illnesses, these potential dangers must be considered in terms of a real risk to public health.

Molecular Detection of Carbapenem Resistance in Acinetobacter Baumannii Isolated From Patients in Khorramabad City, Iran

2020 ◽  
Vol 20 (4) ◽  
pp. 543-549
Author(s):  
Zeinab Babaie ◽  
Somayeh Delfani ◽  
Faranak Rezaei ◽  
Fatemeh Norolahi ◽  
Somayeh Mahdian ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Acinetobacter Baumannii ◽  
Resistance Genes ◽  
High Resistance ◽  
Diffusion Method ◽  
Resistance Pattern ◽  
Biochemical Tests ◽  
Transfer Resistance ◽  
Drug Resistance Pattern ◽  
Wide Range

Background: Acinetobacter baumannii is an opportunistic pathogen, which causes a wide range of infections in hospitals, especially in intensive care units. Nowadays, due to the high resistance of Acinetobacter bumanni to antibiotics, this study, in addition to the phenotypic and genotypic investigations of drug resistance, focused on determining the molecular types of Acinetobacter baumannii isolated from patients in Khorramabad city by the pulsed-field gel electrophoresis (PFGE) method. Materials and Methods: In this cross-sectional study, 50 samples of Acinetobacter baumannii were collected from educational hospitals in Khorramabad city, Iran, from January to August 2015. They were identified in the laboratory using biochemical tests and culture methods. After determining the drug resistance pattern by the disc diffusion method and percentage of resistance genes to carbapenems, Acinetobacter baumannii isolates were analyzed using the PFGE method using the Apa1 enzyme. Results: The highest antibiotic resistance observed for Acinetobacter baumannii strains was against ampicillin-sulbactam (100%) and aztreonam (98%). The highest sensitivity was to polymixin B (100%) and colistin (94%), and also to the OXA-51-like gene present in all samples. The OXA-23-like gene was positive in 44 (88%) samples. PFGE results showed that Acinetobacterbaumannii strains had 33 different pulsotype patterns, of which 27 patterns had more than one strain and 23 had only one strain. Conclusion: Due to the high resistance of Acinetobacter baumannii and its ease of spread and its ability to transfer resistance genes, resistance control methods should be used in the disinfection of hospital areas. Hospital staff should observe hygiene standards and there should also be a reduction in antibiotic use.

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.

Phenotypic detection of extended-spectrum beta-lactamase in multidrug-resistant acinetobacter baumannii isolated in Fauji Foundation Hospital Rawalpindi

2021 ◽  
pp. 1-12
Author(s):  
Asim Ali Shah ◽  
Yasir Ali ◽  
Ayesha Maqbool ◽  
Shahid Ahmad Abbasi ◽  
Admin
Keyword(s):  
Acinetobacter Baumannii ◽  
Sampling Technique ◽  
Polymyxin B ◽  
Multidrug Resistant ◽  
Cross Sectional Study ◽  
Beta Lactamase ◽  
Cross Sectional ◽  
Extended Spectrum Beta Lactamase ◽  
Extended Spectrum ◽  
Multiple Drugs

Abstract Objective: To evaluate the phenotypic detection of extended-spectrum beta-lactamase in multidrug-resistant acinetobacter baumannii. Methods: The cross-sectional study was conducted at the Department of Microbiology, Fauji Foundation Hospital, Rawalpindi, Pakistan, from August 2018 to April 2019, after the ethical approval from the Institutional Review Committee. Consecutive Non- probability sampling technique was used, and comprised clinical specimens, including pus, blood, sputum, urine, tracheal tubes and canula double lumen, which were processed using standard protocols. Colonies of acinetobacter baumannii were identified by gram staining and Analytical Profile Index-20E kit. Combination disc method was used for the identification of extended-spectrum beta-lactamse. Clinical and Laboratory Standards Institute guidelines were used for antimicrobial susceptibility. Data was analysed using SPSS 22 and Sample size was calculated by using earlier study with 5 % margin of error and 95 % confidence level. Results: Of the 78 isolates, 58(74.4%) related to females and 20(25.6%) to males. There was no extended-spectrum beta-lactamse producer. Imipenem, meropenem, cefotaxime, ampicillin and ceftazidime showed 100% resistance, while colistin and polymyxin B were sensitive to all strains. The incidence rate was high in samples isolated from tracheal tubes 47(60.3%), followed by pus 21(26.9%). Age was not found to be a significant factor (p>0.05).   Conclusion: Acinetobacter baumannii showed a high resistance to multiple drugs and was not confined to any specific age group. Colistin and polymyxin B were found to be better choices. Continuous...

scholarly journals Evaluation of antibiotic resistance pattern of Acinetobacter baumannii clinical isolates in Khorramabad hospitals

2021 ◽  
pp. 158-167
Keyword(s):  
Antibiotic Resistance ◽  
Multidrug Resistance ◽  
Acinetobacter Baumannii ◽  
Clinical Isolates ◽  
Clinical Samples ◽  
Resistance Pattern ◽  
Multiple Drug ◽  
Drug Resistant ◽  
Biochemical Tests ◽  
Antibiotic Resistance Pattern

Background: Today, drug-resistant strains of Acinetobacter baumannii are one of the opportunistic pathogens in the world. This study aimed to determine the antibiotic resistance pattern of clinical isolates of A. baumannii in Khorramabad hospitals (i.e., Shohaday Ashayer-Shahid Rahimi), Khorramabad, Iran. Materials and Methods: This cross-sectional study was conducted on the clinical samples collected from patients hospitalized in the different wards of Khorramabad hospitals in 2015-16 and 2017-18. The clinical samples were identified as A. baumannii by microbiological culture and biochemical tests then confirmed by polymerase chain reaction. The susceptibility test of bacterial isolates to antibiotics was performed and the obtained data were analyzed by SPSS software (version 22) using the Chi-square test. Results: According to the results of the antibiogram, among the 94 isolates of A. baumannii collected from the patients admitted to Khorramabad hospitals, 50%, 41.49%, and 8.51% of isolates were multiple-drug resistant, extensively drug-resistant, and non-multidrug resistance, respectively. Conclusion: Due to the sensitivity of A. baumannii to polymyxin B and minocycline antibiotics, these antibiotics can be used, especially as a combination therapy, in the treatment of infections caused by this bacterium. Since the rate of multidrug resistance in Khorramabad hospitals was found to be 91.49%, it is necessary to pay attention to the criteria for controlling nosocomial infections.

scholarly journals Resistance Pattern of Carbapenem on Enterobacteriaceae

2018 ◽  
Vol 56 (214) ◽  
pp. 931-935
Author(s):  
Khilasa Pokharel ◽  
Bishwa Raj Dawadi ◽  
Chandra Prakash Bhatta ◽  
Satish Gupte ◽  
Beena Jha
Keyword(s):  
Culture Media ◽  
Antibiotic Sensitivity ◽  
Polymyxin B ◽  
Control Measures ◽  
Resistance Pattern ◽  
Clinical Microbiology Laboratory ◽  
Cross Sectional ◽  
Carbapenem Resistant ◽  
The Family ◽  
Carbapenem Resistant Enterobacteriaceae

Introduction: Gram negative bacilli are the important causes of common clinical infections. Carbapenem resistant Enterobacteriaceae are considered as important public health threat and is classified as urgent by the Centers of Disease Control and Prevention because of their progressive geographic dissemination and limited therapeutic alternatives. This study was done to find out the resistance pattern of Carbapenem among Enterobacteriaceae.Methods: The descriptive cross-sectional study was carried out in Clinical Microbiology laboratory from February 2018 to May 2018 after ethical approval. Organism was identified on the basis of its microscopic observation by performing Gram’s stain and by identification of morphology after its growth in culture media followed by its biochemical reactions. Antibiotic sensitivity test of isolated pathogens was done using Muller Hinton Agar by the standard disk diffusion technique of Kirby-Bauer method.Results: In our study, total 1055 sample belongs to the family Enterobacteriaceae. From the family Enterobactericeae, 348 (27%) of the bacilli were found to be Carbapenem resistant. Among which most common bacteria was Klebsiella pneumoniae followed by Escherichia coli. All strains of Carbapenem resistant Enterobacteriaceae were sensitive to Colistin, Polymyxin B and Tigecycline. Conclusions: Among Enterobacteriaceae, around one-third of the bacterial isolates were Carbapenem resistant. However, to reduce drug resistance antimicrobial stewardship programme and proper infection control measures is required.

scholarly journals Resistance behaviour of inducible clindamycin-resistant staphylococci from clinical samples and aquatic environments

2014 ◽  
Vol 63 (11) ◽  
pp. 1446-1453 ◽  
Author(s):  
Stefanie Heß ◽  
Claudia Gallert
Keyword(s):  
Species Diversity ◽  
Resistance Genes ◽  
Cross Resistance ◽  
Clinical Samples ◽  
Resistance Pattern ◽  
Aquatic Environments ◽  
Environmental Isolates ◽  
Inducible Clindamycin Resistance ◽  
Genes Encoding ◽  
Resistance Patterns

In this study, the species diversity of staphylococci with inducible resistance to macrolides, lincosamides and streptogramin B (MLSB) isolated from clinical samples, sewage and river water was investigated. Inducible clindamycin resistance was tested using a D-test and macrodilution assays. Inducible cross-resistance (iMLSB phenotype) was examined by PCR of erm gene classes A, B, C, F, G, Q, T and 43. Although ermC was the most frequently detected resistance gene in iMLSB phenotypes of environmental staphylococci (61.2 %), resistance genes encoding iMLSB were more diverse than in staphylococci from hospital samples. In 22.4 % of iMLSB staphylococci from aquatic environments, none of the eight tested erm genes was found. Those isolates and erm43-expressing Staphylococcus lentus displayed low erythromycin MICs (3–16 µg ml−1) compared with ermC-positive environmental staphylococci (≥256 µg ml−1). In contrast to clinical isolates with clearly defined resistance behaviour, resistance patterns against MLSB and MICs for clindamycin of environmental isolates were more diverse. Although the abundance of iMLSB staphylococci in the aquatic environment was lower than in staphylococci from hospital samples, the diversity of resistance genes encoding this phenotype seemed to be higher. Oleandomycin is the best marker to correlate iMLSB phenotype and the respective erm gene. The phenotypical behaviour of environmental isolates may differ from the resistance pattern of clinical iMLSB staphylococci expressing ermA or ermC, and this should be considered for successful treatment of infections.

MOLECULAR CHARACTERIZATION OF GENES ENCODING ACQUIRED CARBAPENEMASE OF CARBAPENEMRESISTANT ACINETOBACTER BAUMANNII ISOLATES

2017 ◽  
pp. 52-57
Author(s):  
Nu Xuan Thanh Le ◽  
Thi Anh Ngoc Le ◽  
Thi Nam Lien Nguyen ◽  
Viet Quynh Tram Ngo ◽  
Santona Antonella ◽  
...  
Keyword(s):  
Acinetobacter Baumannii ◽  
Carbapenem Resistance ◽  
Resistance Rate ◽  
University Hospital ◽  
Resistant Bacteria ◽  
Cross Sectional ◽  
Carbapenem Resistant ◽  
Genes Encoding ◽  
Resistance Rates ◽  
And Control

Background: Today carbapenem-resistant A. baumannii isolates are rising in several parts of the world including Vietnam, they are recognized to be among the most difficult resistant bacteria to treat and control. Carbapenem resistance is most commonly caused by the production of OXA-type carbapenemases and metallo-β-lactamases (MBLs). Objectives: Determine the rate and detect the genes encoding acquired carbapenemase of carbapenem-resistant A. baumannii isolates. Materials and methods: Study design is cross-sectional descriptive study. Carbapenem-resistant A. baumannii isolates in 90 A. baumannii (ACB) complex isolates were collected from Hue Central Hospital (HCH) and Hue University Hospital (HUP). Susceptibility to carbapenem of A. baumannii strains were performed by MicroScan method. Multiplex PCRs were performed to detect the genes encoding acquired carbapenemase. Results: Carbapenem resistance rates in A. baumannii were 88.5% and 87.5% in HCH and HHUMP, respectively. All of genes blaOXA-51, blaOXA-23, blaOXA-58, blaIMP, blaNDM as well as coexistence of two genes (blaIMP, blaNDM) or three genes ((blaOXA-51, blaOXA-23, blaOXA-58) or (blaIMP, blaNDM, blaOXA-58)) were detected in carbapenem resistant A. baumannii isolates. Conclusions: Carbapenem resistance rate in A. baumannii was relatively high. The emergence of carbapenem resistance in A. baumannii is associated with the production of OXA-type carbapenemases and metallo-β-lactamases (MBLs). Key words: Acinetobacter baumannii; carbapenem resistance; carbapenemase

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