Detection of Mycoplasma in Human Cell Cultures Using Homology and PCR

Advisor Information

Paul Davis

Location

UNO Criss Library, Room 231

Presentation Type

Oral Presentation

Start Date

6-3-2015 10:45 AM

End Date

6-3-2015 11:00 AM

Abstract

Mycoplasma is a genus of bacteria which commonly infects not only live humans and other organisms but also the cells of laboratory tissue cultures. Due to their impressively small size, this genus of bacteria is difficult to detect in cell cultures using standard microscopy. Additionally, since Mycoplasma lacks cell walls, they are impervious to common cell-wall disrupting antibiotics often used to treat human cell cultures. Infection of a biological laboratory’s cell cultures with Mycoplasma can disrupt or delay research experiments in a variety of ways, including altering enzyme function, causing chromosomal defects, and slowing cell growth rate. These disruptions can eventually cost a research laboratory both valuable time and money. Therefore, an inexpensive, efficient method to detect these detrimental bacteria would be a wise investment for any biological laboratory to adopt. In this study, we applied one such method for easy detection of Mycoplasma. First, we used the bioinformatics program Clustal to align the genomes of select species of Mycoplasma (those most prone to infect human cell cultures), in order to find homologous genetic sequences among them. Once similar sequences were determined, forward and reverse primers were designed to those sequences to be used in PCR-based detection. Since this method of detection is very specific for strains of Mycoplasma known to infect human cells, and since it is carried out using basic materials found in any biology research laboratory, it is aimed to be both comprehensive and cost-effective.

Additional Information (Optional)

Winner of Outstanding Undergraduate Oral Presentation

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COinS
 
Mar 6th, 10:45 AM Mar 6th, 11:00 AM

Detection of Mycoplasma in Human Cell Cultures Using Homology and PCR

UNO Criss Library, Room 231

Mycoplasma is a genus of bacteria which commonly infects not only live humans and other organisms but also the cells of laboratory tissue cultures. Due to their impressively small size, this genus of bacteria is difficult to detect in cell cultures using standard microscopy. Additionally, since Mycoplasma lacks cell walls, they are impervious to common cell-wall disrupting antibiotics often used to treat human cell cultures. Infection of a biological laboratory’s cell cultures with Mycoplasma can disrupt or delay research experiments in a variety of ways, including altering enzyme function, causing chromosomal defects, and slowing cell growth rate. These disruptions can eventually cost a research laboratory both valuable time and money. Therefore, an inexpensive, efficient method to detect these detrimental bacteria would be a wise investment for any biological laboratory to adopt. In this study, we applied one such method for easy detection of Mycoplasma. First, we used the bioinformatics program Clustal to align the genomes of select species of Mycoplasma (those most prone to infect human cell cultures), in order to find homologous genetic sequences among them. Once similar sequences were determined, forward and reverse primers were designed to those sequences to be used in PCR-based detection. Since this method of detection is very specific for strains of Mycoplasma known to infect human cells, and since it is carried out using basic materials found in any biology research laboratory, it is aimed to be both comprehensive and cost-effective.