Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Mycoplasma detection by PCR analysis

A Hopert1, C C Uphoff, M Wirth

  • 1DSM-German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany.

In Vitro Cellular & Developmental Biology. Animal
|October 1, 1993
PubMed
Summary
This summary is machine-generated.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Mycoplasma hyopneumoniae resides intracellularly within porcine epithelial cells.

Scientific reports·2018
Same author

Cytogenetically cryptic ZMYM2-FLT3 and DIAPH1-PDGFRB gene fusions in myeloid neoplasms with eosinophilia.

Leukemia·2017
Same author

Esterase Isoenzyme Profiles in Acute and Chronic Leukemias.

Leukemia & lymphoma·2016
Same author

Lineage-Specific Monocytic Esterase, a Distinct Marker for Leukemias of Monocytic Origin: Cytochemical, Isoenzymatic and Biochemical Features.

Leukemia & lymphoma·2016
Same author

Acid Phosphatase Isoenzyme Profiles in Acute and Chronic Leukemias.

Leukemia & lymphoma·2016
Same author

Differential expression of TRAP Isoenzyme in B-CLL Cells Treated with Different Inducers.

Leukemia & lymphoma·2016
Same journal

Establishment and characterization of an ovarian cell line from red seabream (Pagrus major) and its application in fish toxicology.

In vitro cellular & developmental biology. Animal·2026
Same journal

DIRAS2 promotes the progression of oral leukoplakia via suppressing ferroptosis.

In vitro cellular & developmental biology. Animal·2026
Same journal

Distinct adhesion energy and extracellular matrix profiles of healthy and osteoarthritic human chondrocytes.

In vitro cellular & developmental biology. Animal·2026
Same journal

Pesticides-induced cellular toxicity in the spinal cord cell line of Lates calcarifer.

In vitro cellular & developmental biology. Animal·2026
Same journal

Serotype-specific effects of adenovirus 5 and 52 E4ORF1 proteins on endothelial cell-mediated regulation of hematopoietic stem cell expansion and lineage differentiation.

In vitro cellular & developmental biology. Animal·2026
Same journal

ALKBH4 confers ferroptosis resistance and drives tumorigenesis via dysregulation of GPX4 in breast cancer cells.

In vitro cellular & developmental biology. Animal·2026
See all related articles

Polymerase chain reaction (PCR) effectively detects mycoplasma contamination in cell lines. This sensitive and specific nested PCR method provides rapid, reliable results for cell culture quality control.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Cell Biology

Background:

  • Mycoplasma contamination is a common issue in continuous cell lines, potentially affecting experimental results.
  • Traditional microbiological cultivation assays can be time-consuming and may lack sensitivity.

Purpose of the Study:

  • To evaluate the efficacy of a nested polymerase chain reaction (PCR) assay for detecting mycoplasma contamination in cell lines.
  • To compare the PCR method with traditional agar-based cultivation assays.

Main Methods:

  • A nested, double-step PCR assay using primers targeting conserved regions of the 16S rRNA gene was developed.
  • The PCR assay was applied to a panel of 42 continuous cell lines, with results compared to agar cultivation.

Main Results:

Related Experiment Videos

  • The PCR assay demonstrated high sensitivity and specificity, accurately identifying all contaminated and uncontaminated cell lines.
  • Distinctive DNA bands were observed in PCR-amplified samples from contaminated cell lines, with no cross-hybridization to eukaryotic DNA.
  • The PCR method yielded no false-positive or false-negative results when proper conditions were maintained.

Conclusions:

  • Nested PCR is a highly sensitive, specific, reliable, and rapid method for detecting mycoplasma contamination in cell cultures.
  • This PCR assay offers significant advantages over traditional cultivation methods for routine cell line screening.
  • The method ensures accurate cell culture quality control, preventing compromised research outcomes.