Validation of a PCR coupled to a microarray method for detection of mycoplasma in vaccines

Biologicals. 2017 Nov:50:55-62. doi: 10.1016/j.biologicals.2017.09.001. Epub 2017 Sep 23.

Abstract

The revised section of the European, United States, and Japan Pharmacopeias on mycoplasma testing provided guidance for the set up and validation of a nucleic acid amplification technique (NAT) as an alternative method to agar culture and indicator cell culture compendial methods. The CytoInspect™ method, based on Polymerase Chain Reaction (PCR) coupled to microarray analysis, has been selected for detection and identification of mycoplasma in vaccines. To replace compendial methods, the alternative method must demonstrate equivalence in both limit of detection (LOD) and specificity compared with compendial methods. Here, we summarize the validation of the CytoInspect™ method according to current pharmacopeia requirements. Validation of the robustness, sensitivity (at least 10 colony forming units/ml) and specificity of the CytoInspect™ method are demonstrated. Likewise, a comparability study was performed to compare the LOD for CytoInspect™ compared with the previously validated LOD for compendial culture tests.

Keywords: European; Mycoplasma; PCR coupled to microarray; United States and Japan pharmacopeias; Validation.

Publication types

  • Validation Study

MeSH terms

  • Animals
  • Cell Line
  • Chlorocebus aethiops
  • Guidelines as Topic / standards
  • Humans
  • Limit of Detection
  • Microarray Analysis / methods
  • Mycoplasma / genetics
  • Mycoplasma / isolation & purification*
  • Mycoplasma Infections / microbiology*
  • Pharmacopoeias as Topic / standards
  • Polymerase Chain Reaction / methods*
  • Reproducibility of Results
  • Vaccines / standards*
  • Vero Cells

Substances

  • Vaccines