---
title: June 2022 Update to the PathoSEEK® Total Yeast and Mold Method
description: On June 16th, 2022 MGC released an updated user guide and validation for the Total Yeast and Mold method.
---
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# June 2022 Update to the PathoSEEK® Total Yeast and Mold Method
## On June 16th, 2022 MGC released an updated user guide and validation for the Total Yeast and Mold method.
This update is a result of months of torture testing our TYM process, during which:
- Six different internal lab users tested the same samples to understand the process variance
- A beta-testing site ran the updated protocol on real-life samples and achieved concordance with plating
- Inclusion and exclusion testing was expanded to 30 and 50 organisms respectively as seen in other AOAC SMPRs
[Download the PathoSEEK® Total Yeast and Mold Detection Assay with SenSATIVAx® & TLP Extraction Validation Document](https://help.medicinalgenomics.com/hubfs/Validation%20Documents/Total%20Yeast%20and%20Mold%20with%20SenSATIVAx%C2%AE%20TLP%20%20Validation%20Document.pdf?utm_campaign=Support%20Communication&utm_source=hs_email&utm_medium=email&_hsenc=p2ANqtz--olSCzfVIOUlHrltJLWsE2rgvWLahYQWcrcDvg7H8z1C4IECvOHVjjfua2PvppBlKA3Y25&hsLang=en)
**New PT Recommendation for flower**
In order for your lab to validate the new PathoSEEK® Total Yeast & Mold Detection Assay with SenSATIVAx® Extraction protocol, we recommend the following standard from NSI:
- PT Express Catalog Number CMPT-085B from NSI Lab Solutions
This new CRM includes both yeast and mold organisms, which more accurately represents what labs encounter on real-life samples and achieves better concordance when comparing qPCR and plating results.
**Important information regarding Cq to CFU conversion equations for TYM Testing:**
The Medicinal Genomics Total Yeast and Mold assay targets ITS (Internal Transcribed Spacer) regions in the yeast and filamentous fungi genomes. The prevalence of these regions can vary by 10-100x between organisms.
Mathematically, The equation assumes 1 genomic copy = 1 PCR target. Yeast have ~20 ITS regions per genome and molds have 50-100 ITS regions which means when amplifying these organisms in situ, you’re not amplifying 1 copy, but 20-100 copies per copy of the genome. This increases the sensitivity of qPCR when targeting a region present in multiple copies per genome.
Plate enumerations for yeast and mold is highly variable due to 4 variables involved that can vastly affect results
1. Time the plate is incubated
2. Carbon source
3. Temperature used
4. Organisms that do not culture
Using an assay that targets a genomic region present in 20-100 copies is less variable than juggling the 4 previously mentioned growth factors. In addition, having 10X higher sensitivity or 100X higher sensitivity isn't a deficit when you are comparing it to a ‘gold standard’ of plating which also accepts that many microbes are unculturable.
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