PCR-BASED DETECTION OF MYCOPLASMA SPP. CONTAMINATION IN CELL CULTURE

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SHAHHOSSEINY M.H.; HOSSEINY Z.; AKHLAGHI F.; SHOKRGOZAR M.A.

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Journal: JOURNAL OF MICROBIOLOGY KNOWLEDGE Year:2010 | Volume:2 | Issue:6 Page(s): 13-25

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Title

PCR-BASED DETECTION OF MYCOPLASMA SPP. CONTAMINATION IN CELL CULTURE

Author(s)

SHAHHOSSEINY M.H. | HOSSEINY Z. | AKHLAGHI F. | SHOKRGOZAR M.A. | Issue Writer Certificate

Keywords

MYCOPLASMAQ3

PCRQ4

CONTAMINATIONQ3

MOLECULAR DIAGNOSISQ3

CELL CULTUREQ3

Abstract

Infections with MYCOPLASMA species can induce a variety of problems in living organisms and in in vitro CELL CULTUREs. These non-apparent infections tend to be quite insidious, as they may affect a variety of biochemical and genetic aspects of the infected cells, thereby resulting in unreliable experimental results and the possible transmission of diseases. Therefore, it is necessary to establish a routine diagnostic protocol for MYCOPLASMA infection in order to ensure reliable research results, as well as the safety of commercial biological products. However, the detection of Mycoplasmic species in CELL CULTUREs and another biological product remains a problem. The majority of currently available detection procedures are not sufficient for the simultaneous detection of the major MYCOPLASMA species contaminants commonly encountered in in vitro CELL CULTUREs. In order to circumvent those limitations, many nucleic acid technology-predicated procedures have been developed. PCR-based methods for the detection of certain DNA regions of the MYCOPLASMA genome have proven both rapid and specific. Using SHAH-GPO-3, MGSO primers and standard MYCOPLASMA species PCR optimized and sensitivity and specificity evaluated by Known CFU samples and different strains. CELL CULTURE DNA extracted and then tested by optimized PCR. Amplicon (272 bp) cloned by PCR-cloning and then sequenced by di-deoxy chain termination. In this study, we describe our newly-developed sensitive PCR procedure for the detection of MYCOPLASMA genus contaminants. For amplification, the DNA regions of 16SrRNA were targeted using general MYCOPLASMA primers. The PCR, which generated DNA fragments of 272 bp, was found to be able to detect 10 copies of the target DNA, and evidenced no cross-reactivity with the genomic DNA of related microorganisms or of human cell lines, thereby confirming the sensitivity and specificity of the primers used. 25 from 47 cell lines infected with MYCOPLASMA. The results obtained in this study furnish evidence suggesting that the employed assay system constitutes an effective tool for the diagnosis of MYCOPLASMAl CONTAMINATION in CELL CULTURE and another biological system.

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APA: Copy

SHAHHOSSEINY, M.H., HOSSEINY, Z., AKHLAGHI, F., & SHOKRGOZAR, M.A.. (2010). PCR-BASED DETECTION OF MYCOPLASMA SPP. CONTAMINATION IN CELL CULTURE. JOURNAL OF MICROBIOLOGY KNOWLEDGE, 2(6), 13-25. SID. https://sid.ir/paper/193205/en

Vancouver: Copy

SHAHHOSSEINY M.H., HOSSEINY Z., AKHLAGHI F., SHOKRGOZAR M.A.. PCR-BASED DETECTION OF MYCOPLASMA SPP. CONTAMINATION IN CELL CULTURE. JOURNAL OF MICROBIOLOGY KNOWLEDGE[Internet]. 2010;2(6):13-25. Available from: https://sid.ir/paper/193205/en

IEEE: Copy

M.H. SHAHHOSSEINY, Z. HOSSEINY, F. AKHLAGHI, and M.A. SHOKRGOZAR, “PCR-BASED DETECTION OF MYCOPLASMA SPP. CONTAMINATION IN CELL CULTURE,” JOURNAL OF MICROBIOLOGY KNOWLEDGE, vol. 2, no. 6, pp. 13–25, 2010, [Online]. Available: https://sid.ir/paper/193205/en

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