2010_Rapid Detection and Identification of Methicillin-Resistant Staphylococcus Aureus (MRSA) By Real-Time Polymerase Chain Reaction (RT-PCR)
| Format: | General Document |
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| _version_ | 1860798011236417536 |
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| building | INTELEK Repository |
| collection | Online Access |
| collectionurl | https://intelek.unisza.edu.my/intelek/pages/search.php?search=!collection3 |
| copyright | Copyright©PWB2025 |
| country | Malaysia |
| date | 2012-02-21 |
| format | General Document |
| id | 15436 |
| institution | UniSZA |
| internalnotes | Sila masukkan subject wajib Dissertations, Academic untuk semua tesis.. Terima kasih |
| originalfilename | RAPID DETECTION AND IDENTIFICATION OF METHICILLIN-RESISTANT Staphylococcus aureus (MRSA) BY REAL-TIME POLYMERASE CHAIN REACTION (RT-PCR) (MASTER_2010).pdf |
| person | Zarizal Bin Suhaili |
| recordtype | oai_dc |
| resourceurl | https://intelek.unisza.edu.my/intelek/pages/view.php?ref=15436 |
| sourcemedia | Server storage Scanned document |
| spelling | 15436 https://intelek.unisza.edu.my/intelek/pages/view.php?ref=15436 https://intelek.unisza.edu.my/intelek/pages/search.php?search=!collection3 General Document Malaysia Library Staff (Top Management) Library Staff (Management) Library Staff (Support) Terengganu Faculty of Bio-resources & Food Industry English application/pdf 1.5 142 Server storage Scanned document Universiti Sultan Zainal Abidin UniSZA Private Access UNIVERSITI SULTAN ZAINAL ABIDIN SAMBox 2.3.4; modified using iTextSharp™ 5.5.10 ©2000-2016 iText Group NV (AGPL-version) RAPID DETECTION AND IDENTIFICATION OF METHICILLIN-RESISTANT Staphylococcus aureus (MRSA) BY REAL-TIME POLYMERASE CHAIN REACTION (RT-PCR) (MASTER_2010).pdf 2010_Rapid Detection and Identification of Methicillin-Resistant Staphylococcus Aureus (MRSA) By Real-Time Polymerase Chain Reaction (RT-PCR) Zarizal Bin Suhaili 2012-02-21 Copyright©PWB2025 Methicillin-resistant Staphylococcus aureus Methicillin-resistance Staphylococcus aureus (MRSA) has been reported as one of the major contributor of nosocomial infection in the world and ranks as one of the most difficult pathogen to treat in hospitalized patients. This study aimed to utilize molecular diagnostic techniques for the detection and identification of Staphylococcus aureus especially MRSA as well as methicillin-sensitive Staphylococcus aureus (MSSA) and subsequently to study antibiotic biograms profiles of selected isolates. Thirty-two clinical isolates of S. aureus from two teaching hospitals, University Malaya Medical Center (UMMC), Hospital University of Kebangsaan Malaysia (HUKM) and Hospital Sultanah Nur Zahirah (HSNZ), Kuala Terengganu were used in this study. The antibiogram profile of S. aureus clinical isolates from HSNZ and reference isolates form American Type Culture Collection (ATCC) was established by using agar disc diffusions methods. Out of the 19 isolates from HSNZ, 11 were confirmed as MRSA which exhibiting multidrug resistance against amikacin, cefoxitin, gentamycin, clindamycin, erythromycin, fusidic acid, norfloxacin, oxacilin, penicillin, rifampin, streptomycin, tetracycline and trimetroprim. All isolates were still susceptible to vancomycin. Apart from vancomycin, four other antibiotics used i.e., chloramphenicol, linezolid, novobiocin and teicoplanin seem to be the most effective antibiotic against all S. aureus isolates. Rapid molecular diagnostic by real-time PCR SYBR Green I was established. Development and verification of real-time PCR primers by in-silico PCR was established. Three selected genes as a species-specific marker coa (staphylocoaguase), nuc (thermonuclease) and mecA (methicillin-reistance) were verified against 18 selected S. aureus strains by using in-silico PCR. The actual laboratory verification, of all three selected genes were positively produced single specific melting curve analysis peak Tm at 76.16 ± 0.8 °C, 78.5 ± 0.4 °C and 74.41 ± 0.6 °C for coa, nuc, and mecA respectively against 32 bacterial strains including ATCC reference strains. There was no disagreement between in-silico PCR and real time PCR. In this study a MRSA detection method based on the melting temperature analysis profiling was established. Simplex and duplex real-time PCR assay were used for the simultaneous detection of nuc (species-specific) and mecA (methicillin Methicillin-resistance Staphylococcus aureus (MRSA) has been reported as one of the major contributor of nosocomial infection in the world and ranks as one of the most difficult pathogen to treat in hospitalized patients. This study aimed to utilize molecular diagnostic techniques for the detection and identification of Staphylococcus aureus especially MRSA as well as methicillin-sensitive Staphylococcus aureus (MSSA) and subsequently to study antibiotic biograms profiles of selected isolates. Thirty-two clinical isolates of S. aureus from two teaching hospitals, University Malaya Medical Center (UMMC), Hospital University of Kebangsaan Malaysia (HUKM) and Hospital Sultanah Nur Zahirah (HSNZ), Kuala Terengganu were used in this study. The antibiogram profile of S. aureus clinical isolates from HSNZ and reference isolates form American Type Culture Collection (ATCC) was established by using agar disc diffusions methods. Out of the 19 isolates from HSNZ, 11 were confirmed as MRSA which exhibiting multidrug resistance against amikacin, cefoxitin, gentamycin, clindamycin, erythromycin, fusidic acid, norfloxacin, oxacilin, penicillin, rifampin, streptomycin, tetracycline and trimetroprim. All isolates were still susceptible to vancomycin. Apart from vancomycin, four other antibiotics used i.e., chloramphenicol, linezolid, novobiocin and teicoplanin seem to be the most effective antibiotic against all S. aureus isolates. Rapid molecular diagnostic by real-time PCR SYBR Green I was established. Development and verification of real-time PCR primers by in-silico PCR was established. Three selected genes as a species-specific marker coa (staphylocoaguase), nuc (thermonuclease) and mecA (methicillin-reistance) were verified against 18 selected S. aureus strains by using in-silico PCR. The actual laboratory verification, of all three selected genes were positively produced single specific melting curve analysis peak Tm at 76.16 ± 0.8 °C, 78.5 ± 0.4 °C and 74.41 ± 0.6 °C for coa, nuc, and mecA respectively against 32 bacterial strains including ATCC reference strains. There was no disagreement between in-silico PCR and real time PCR. In this study a MRSA detection method based on the melting temperature analysis profiling was established. Simplex and duplex real-time PCR assay were used for the simultaneous detection of nuc (species-specific) and mecA (methicillin-resistance) genes in a single SYBR Green I real-time PCR tube assay. Evaluations were based on the melting temperature (Tm) analysis of the amplicons using 23 S. aureus clinical isolates including three ATCC S. aureus standard strains. Real-time PCR amplification products with melting peaks at 78.39 ± 0.4 ºC and 74.41 ± 0.6 ºC were detected for nuc and mecA genes, respectively. Each real-time PCR assay was completed within two hours. This rapid genotypic method is useful for the detection of resistant determinant (mecA) and identification of S. aureus (nuc) clinical isolates. Dissertations, Academic Sila masukkan subject wajib Dissertations, Academic untuk semua tesis.. Terima kasih Methicillin-Resistant Staphylococcus Aureus (MRSA) Real-time Polymerase Chain Reaction (RT-PCR) Rapid Detection Thesis |
| spellingShingle | 2010_Rapid Detection and Identification of Methicillin-Resistant Staphylococcus Aureus (MRSA) By Real-Time Polymerase Chain Reaction (RT-PCR) |
| state | Terengganu |
| subject | Methicillin-resistant Staphylococcus aureus Dissertations, Academic |
| summary | Methicillin-resistance Staphylococcus aureus (MRSA) has been reported as one of the major contributor of nosocomial infection in the world and ranks as one of the most difficult pathogen to treat in hospitalized patients. This study aimed to utilize molecular diagnostic techniques for the detection and identification of Staphylococcus aureus especially MRSA as well as methicillin-sensitive Staphylococcus aureus (MSSA) and subsequently to study antibiotic biograms profiles of selected isolates. Thirty-two clinical isolates of S. aureus from two teaching hospitals, University Malaya Medical Center (UMMC), Hospital University of Kebangsaan Malaysia (HUKM) and Hospital Sultanah Nur Zahirah (HSNZ), Kuala Terengganu were used in this study. The antibiogram profile of S. aureus clinical isolates from HSNZ and reference isolates form American Type Culture Collection (ATCC) was established by using agar disc diffusions methods. Out of the 19 isolates from HSNZ, 11 were confirmed as MRSA which exhibiting multidrug resistance against amikacin, cefoxitin, gentamycin, clindamycin, erythromycin, fusidic acid, norfloxacin, oxacilin, penicillin, rifampin, streptomycin, tetracycline and trimetroprim. All isolates were still susceptible to vancomycin. Apart from vancomycin, four other antibiotics used i.e., chloramphenicol, linezolid, novobiocin and teicoplanin seem to be the most effective antibiotic against all S. aureus isolates. Rapid molecular diagnostic by real-time PCR SYBR Green I was established. Development and verification of real-time PCR primers by in-silico PCR was established. Three selected genes as a species-specific marker coa (staphylocoaguase), nuc (thermonuclease) and mecA (methicillin-reistance) were verified against 18 selected S. aureus strains by using in-silico PCR. The actual laboratory verification, of all three selected genes were positively produced single specific melting curve analysis peak Tm at 76.16 ± 0.8 °C, 78.5 ± 0.4 °C and 74.41 ± 0.6 °C for coa, nuc, and mecA respectively against 32 bacterial strains including ATCC reference strains. There was no disagreement between in-silico PCR and real time PCR. In this study a MRSA detection method based on the melting temperature analysis profiling was established. Simplex and duplex real-time PCR assay were used for the simultaneous detection of nuc (species-specific) and mecA (methicillin Methicillin-resistance Staphylococcus aureus (MRSA) has been reported as one of the major contributor of nosocomial infection in the world and ranks as one of the most difficult pathogen to treat in hospitalized patients. This study aimed to utilize molecular diagnostic techniques for the detection and identification of Staphylococcus aureus especially MRSA as well as methicillin-sensitive Staphylococcus aureus (MSSA) and subsequently to study antibiotic biograms profiles of selected isolates. Thirty-two clinical isolates of S. aureus from two teaching hospitals, University Malaya Medical Center (UMMC), Hospital University of Kebangsaan Malaysia (HUKM) and Hospital Sultanah Nur Zahirah (HSNZ), Kuala Terengganu were used in this study. The antibiogram profile of S. aureus clinical isolates from HSNZ and reference isolates form American Type Culture Collection (ATCC) was established by using agar disc diffusions methods. Out of the 19 isolates from HSNZ, 11 were confirmed as MRSA which exhibiting multidrug resistance against amikacin, cefoxitin, gentamycin, clindamycin, erythromycin, fusidic acid, norfloxacin, oxacilin, penicillin, rifampin, streptomycin, tetracycline and trimetroprim. All isolates were still susceptible to vancomycin. Apart from vancomycin, four other antibiotics used i.e., chloramphenicol, linezolid, novobiocin and teicoplanin seem to be the most effective antibiotic against all S. aureus isolates. Rapid molecular diagnostic by real-time PCR SYBR Green I was established. Development and verification of real-time PCR primers by in-silico PCR was established. Three selected genes as a species-specific marker coa (staphylocoaguase), nuc (thermonuclease) and mecA (methicillin-reistance) were verified against 18 selected S. aureus strains by using in-silico PCR. The actual laboratory verification, of all three selected genes were positively produced single specific melting curve analysis peak Tm at 76.16 ± 0.8 °C, 78.5 ± 0.4 °C and 74.41 ± 0.6 °C for coa, nuc, and mecA respectively against 32 bacterial strains including ATCC reference strains. There was no disagreement between in-silico PCR and real time PCR. In this study a MRSA detection method based on the melting temperature analysis profiling was established. Simplex and duplex real-time PCR assay were used for the simultaneous detection of nuc (species-specific) and mecA (methicillin-resistance) genes in a single SYBR Green I real-time PCR tube assay. Evaluations were based on the melting temperature (Tm) analysis of the amplicons using 23 S. aureus clinical isolates including three ATCC S. aureus standard strains. Real-time PCR amplification products with melting peaks at 78.39 ± 0.4 ºC and 74.41 ± 0.6 ºC were detected for nuc and mecA genes, respectively. Each real-time PCR assay was completed within two hours. This rapid genotypic method is useful for the detection of resistant determinant (mecA) and identification of S. aureus (nuc) clinical isolates. |
| title | 2010_Rapid Detection and Identification of Methicillin-Resistant Staphylococcus Aureus (MRSA) By Real-Time Polymerase Chain Reaction (RT-PCR) |
| title_full | 2010_Rapid Detection and Identification of Methicillin-Resistant Staphylococcus Aureus (MRSA) By Real-Time Polymerase Chain Reaction (RT-PCR) |
| title_fullStr | 2010_Rapid Detection and Identification of Methicillin-Resistant Staphylococcus Aureus (MRSA) By Real-Time Polymerase Chain Reaction (RT-PCR) |
| title_full_unstemmed | 2010_Rapid Detection and Identification of Methicillin-Resistant Staphylococcus Aureus (MRSA) By Real-Time Polymerase Chain Reaction (RT-PCR) |
| title_short | 2010_Rapid Detection and Identification of Methicillin-Resistant Staphylococcus Aureus (MRSA) By Real-Time Polymerase Chain Reaction (RT-PCR) |
| title_sort | 2010_rapid detection and identification of methicillin-resistant staphylococcus aureus (mrsa) by real-time polymerase chain reaction (rt-pcr) |