Determination of PCR-ELISA Diagnostic Value in Comparison With Classical Methods and PCR to Detect Resistance to Methacillin

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Rezavand Susan; AMANI JAFAR; AKBARI NEDA; MOHAJERANI HAMID REZA; NAZARIAN SHAHRAM; MAHMOODZADEH HOSSEINI HAMIDEH; MOOSAZADEH MOGHADDAM MEHRDAD

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Journal: Iranian Journal of Medical Microbiology Year:2019 | Volume:13 | Issue:1 Page(s): 22-31

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Information Journal Paper

Title

Determination of PCR-ELISA Diagnostic Value in Comparison With Classical Methods and PCR to Detect Resistance to Methacillin

Author(s)

Keywords

Staphylococcus aureusQ2

Antibiotic resistanceQ3

PCR-ELISAQ1

mecA geneQ3

Abstract

Background and Aims: High prevalence of Methicillin Resistant Staphylococcus aureus isolates (MRSA) as well as the multi-drug resistance in this bacterium causes difficulties in the treatment of infections due to these bacteria. Hence, detection of MRSA isolates by rapid and accurate methods is necessary. PCR-ELISA is an accurate and molecular technique that is used for the detection of several pathogens. The aim of this study is the detection of MRSA using PCRELISA. Materials and Methods: Specific primers for mecA gene were designed. Then, dNTP labeled with Digoxigenin was applied for amplifying mecA gene. DIG-labeled PCR products were seeded on the well coated streptoavidin and identified by anti-DIG– peroxidase conjugate. Furthermore, Biotin-labeled DNA probe specific for mecA gene was used. Sensitivity and specificity of the method was determined. Resistance to methicillin among 70 clinical isolates was determined by the disk diffusion, agar dilution and PCR-ELISA methods. Results: mecA gene of S. aureus was amplified using gene specific primers resulted in a fragment with 310 bp length. Findings from the PCR-ELISA technic showed no cross-reactivity with Klebsiella Pneumoniae, Bacillus subtilis and Esheriashia coli as control bacteria and its sensitivity was 0. 5 ng. The prevalence of MRSA clinical isolates by the disk diffusion, agar dilution and PCR-ELISA methods was 60%, 58. 5% and 60%, respectively. Conclusion: The PCR-ELISA technique was known as an accurate and rapid test for the detection of infection agents using their specific gene. This technic can applied as an appropriate alternative method for time-consuming, less sensitive and expensive techniques such as Real-time PCR and differential biochemical tests which are currently used in laboratory.

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References

Prevalence and Genotyping of Methicillin-Resistant and-Susceptible Staphylococcus aureus Strains Isolated from Patients in a University Hospital, Isfahan, Iran

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APA: Copy

Rezavand, Susan, AMANI, JAFAR, AKBARI, NEDA, MOHAJERANI, HAMID REZA, NAZARIAN, SHAHRAM, MAHMOODZADEH HOSSEINI, HAMIDEH, & MOOSAZADEH MOGHADDAM, MEHRDAD. (2019). Determination of PCR-ELISA Diagnostic Value in Comparison With Classical Methods and PCR to Detect Resistance to Methacillin. IRANIAN JOURNAL OF MEDICAL MICROBIOLOGY, 13(1 ), 22-31. SID. https://sid.ir/paper/366245/en

Vancouver: Copy

Rezavand Susan, AMANI JAFAR, AKBARI NEDA, MOHAJERANI HAMID REZA, NAZARIAN SHAHRAM, MAHMOODZADEH HOSSEINI HAMIDEH, MOOSAZADEH MOGHADDAM MEHRDAD. Determination of PCR-ELISA Diagnostic Value in Comparison With Classical Methods and PCR to Detect Resistance to Methacillin. IRANIAN JOURNAL OF MEDICAL MICROBIOLOGY[Internet]. 2019;13(1 ):22-31. Available from: https://sid.ir/paper/366245/en

IEEE: Copy

Susan Rezavand, JAFAR AMANI, NEDA AKBARI, HAMID REZA MOHAJERANI, SHAHRAM NAZARIAN, HAMIDEH MAHMOODZADEH HOSSEINI, and MEHRDAD MOOSAZADEH MOGHADDAM, “Determination of PCR-ELISA Diagnostic Value in Comparison With Classical Methods and PCR to Detect Resistance to Methacillin,” IRANIAN JOURNAL OF MEDICAL MICROBIOLOGY, vol. 13, no. 1 , pp. 22–31, 2019, [Online]. Available: https://sid.ir/paper/366245/en

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