MycoQSearch Mycoplasma qPCR Detection Kit

CHARACTERISTICS OF CELLSAFE’S MYCOPLASMA DETECTION KIT

CellSafe’s Mycoplasma Detection kits utilize the PCR technique which is established as the method of choice for the highest sensitivity in the detection of mycoplasma contamination in cell culture and other cell culture derived biologicals. Detection of M.orale, M.hyorhinis, M.arginini, M.fermentans, M.hominis and Acholeplasma laidlawii usually encountered as contaminants in cell cultures. Furthermore, these kits can detect M.pneumoniae, M.salivarium, M.genitalium, M.penentrans and Ureaplasma species. The primers are specific to the highly conserved 16s rRNA coding region and the kits do not detect other bacteria and cell-lines. All kits include Internal Amplification Control (IAC) providing validity of test results. They are user firendly, ready-to-use, optimized PCR master mix type kits.

MycoQSearch^TM Mycoplasma qPCR Detection Kit

The MycoQSearch^TM Mycoplasma qPCR Detection Kit is used to detect Mycoplasma infection of cell cultures by real-time quantitative PCR (qPCR) using Probe. The MycoQSearch^TM Mycoplasma qPCR Detection Kit includes a Primer and Probe mixes. These mixes contain FAM labeled probe specific for mycoplasma species and Hex labeled probe for internal control DNA. The primer set is specific to the highly conserved the 16S rRNA coding region in the mycoplasma genome. This allows the detection of M. orale, M. hyorhinis, M. arginini, M. fermentans, Acholeplasma laidlawii, M. hominis, usually encountered as contaminants in cell cultures. Furthermore, this kit can detect M. pneumoniae, M. salivarium, M. synoviae and Ureaplasma species. Eukaryotic and bacterial DNA is not amplified by MycoQsearch^TM Mycoplasma qPCR Detection Kit.

The MycoQSearch^TM Mycoplasma qPCR Detection Kit is capable of detecting Mycoplasma infections in cell cultures in less than three hours, depending on the spectrofluorometric thermal cycler used for detection.

Kit Specificity

The MycoQSearch^TM Mycoplasma qPCR Detection Kit detects Mycoplasma species simply, reliably, and rapidly. To detect the presence of these microorganisms, the assay uses the polymerase chain reaction (PCR) to amplify a target unique to a wide variety of mycoplasmas. The kit can detect almost all kinds of Mycoplasma species, including Acholeplasma laidlawii and Spiroplasma citri. The kit does not detect other genera or cellline DNA.

Kit Sensitivity

The sensitivity of the PCR using this kit is 1 to 10 copies of the target DNA per reaction. Sensitivity of the assay in real culture samples depends on the quality of the sample preparation.

Results

A successfully performed PCR without inhibition is indicated by an increasing fluorescence signal in the internal control channel (HEX). The internal DNA can be detected with a yellow filter (535-555 nm for HEX). The presence of mycoplasma DNA in the sample is indicated by an increasing fluorescence signal at 510 nm (FAM) and is usually detected with a green filter (470-510 nm).

False-negative results (due to inhibition of PCR reaction by the sample matrix) can be detected individualy for each sample as these reactions do not show any fluorescence signal.  

Internal Control Amplification – HEX (Yellow channel)                   Sample and Positive Control Amplification – FAM (Green channel)