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  1. Laboratory Methods (Food)

BAM Chapter 19b: Molecular Detection of Cyclospora cayetanensis in Fresh Produce Using Real-Time PCR

Bacteriological Analytical Manual (BAM) Main Page

Authors

Sonia Almeria, John Grocholl, Mauricio Durigan, Laura Ewing-Peeples

Contact: Sonia Almeria or Mauricio Durigan

Editions 

Current Edition

Previous Editions 

Revision History

  • May 2026: A matrix extension was completed for detection of C. cayetanensis on fresh basil and parsley and approved by the Microbiology Methods Validation Subcommittee in March 2026. 
  • December 2024: Modification for the use of a new qPCR target (Mit1C).  The previous real-time PCR detection reference method in BAM Chapter 19b (Edition 2022) targeted the C. cayetanensis multicopy 18S ribosomal RNA gene. A more specific qPCR target (Mit1C qPCR) replaces the previous 18S qPCR method to be used by FDA regulatory laboratories.
  • April 2022: C. cayetanensis Blueberry Extension Report has been added.
  • April 2022: Method Modification Due to the Discontinuation of the Current/Original QuantiFast Multiplex PCR kit and Replacement with TaqMan™ Fast Advanced Master Mix Verification Study has been added.
  • December 2020: Cyclospora cayetanensis Blackberry and Shredded Cabbage Reports have been added.
  • November 2019: Cyclospora cayetanensis Romaine lettuce Extension Report has been added.
  • September 2017: Modified FastDNA Spin Extraction Protocol section: Duplicate text removed from step C.; The word "shaking" has been replaced with the word "inverting" in step G.
  • September 2017: Posted a Published Journal Article (PDF) and a Supplemental Data File (PDF).
  • August 2017: Cyclospora cayetanensis Basil Extension Report, C. cayetanensis Parsley Extension Report, and C. cayetanensis Carrot Extension Report have been added.
  • Six Appendices (PDF format) are available at the end of this method.
  • New BAM Chapter 19b; June 2017: Replaces all aspects of the Cyclospora methodology in BAM Chapter 19a related to detection of C. cayetanensis in produce.
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