Interlaboratory assays from the fungal PCR Initiative and the Modimucor Study Group to improve qPCR detection of Mucorales DNA in serum: one more step toward standardization

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F65269705%3A_____%2F25%3A00081021" target="_blank" >RIV/65269705:_____/25:00081021 - isvavai.cz</a>

Result on the web

<a href="https://journals.asm.org/doi/full/10.1128/jcm.01525-24" target="_blank" >https://journals.asm.org/doi/full/10.1128/jcm.01525-24</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1128/jcm.01525-24" target="_blank" >10.1128/jcm.01525-24</a>

Result language

angličtina

Original language name

Interlaboratory assays from the fungal PCR Initiative and the Modimucor Study Group to improve qPCR detection of Mucorales DNA in serum: one more step toward standardization

Original language description

The aim of this study was to identify parameters influencing DNA extraction and PCR amplification efficiencies in an attempt to standardize Mucorales qPCR. The Fungal PCR Initiative Mucorales Laboratory Working Group distributed two panels of simulated samples to 26 laboratories: Panel A (six sera spiked with Mucorales DNA and one negative control serum) and Panel B (six Mucorales DNA extracts). Panel A underwent DNA extraction in each laboratory according to the local procedure and were sent to a central laboratory for testing using three different qPCR techniques: one in-house qPCR assay and two commercial assays (MucorGenius and Fungiplex). Panel B DNA extracts were PCR amplified in each laboratory using local procedures: nine in-house qPCR assays and two commercial kits (MucorGenius and MycoGENIE). All data were compiled and anonymously analyzed at the central laboratory. For Panel A, a total of six different automated platforms and five manual extraction methods were used. Positive rates were 64%, 70%, and 89%, for the MucorGenius, Fungiplex, and the in-house qPCR assay, respectively. Using a large volume of serum for DNA extraction provided the highest analytical sensitivity (82.5% for 1 mL compared with 62.7% for smaller volumes, P &lt; 0.01). For Panel B, five in-house qPCR assays and two commercial kits had &gt;78% positivity. Using larger PCR input volumes (&gt;= 7 mu L) was associated with the highest sensitivity at 95.5% compared to 58.3% when lower input volumes were used (P &lt; 0.01). Using larger sample volumes for nucleic acid extraction and DNA template volumes for PCR amplification significantly improves the performance of Mucorales qPCR when testing serum.

Czech name

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Czech description

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Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

—

OECD FORD branch

10606 - Microbiology

Project

—

Continuities

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Publication year

2025

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

Journal of Clinical Microbiology

ISSN

0095-1137

e-ISSN

1098-660X

Volume of the periodical

63

Issue of the periodical within the volume

2

Country of publishing house

US - UNITED STATES

Number of pages

15

Pages from-to

nestránkováno

UT code for WoS article

001386374600001

EID of the result in the Scopus database

2-s2.0-85219028915