Development of a Novel Multiplex Multiple Oligonucleotide
Ligation PCR System for Identification of Probiotic Bacteria in
Food Supplements

J 2025

Development of a Novel Multiplex Multiple Oligonucleotide Ligation PCR System for Identification of Probiotic Bacteria in Food Supplements

KRÁLÍK, Petr; Martin KLANICA a Radka DZIEDZINSKÁ

Základní údaje

Originální název

Development of a Novel Multiplex Multiple Oligonucleotide Ligation PCR System for Identification of Probiotic Bacteria in Food Supplements

Autoři

KRÁLÍK, Petr; Martin KLANICA a Radka DZIEDZINSKÁ

Vydání

JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, WASHINGTON, AMER CHEMICAL SOC, 2025, 0021-8561

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10606 Microbiology

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 6.200 v roce 2024

Označené pro přenos do RIV

Ano

Kód RIV

RIV/00216224:14310/25:00143442

Organizační jednotka

Přírodovědecká fakulta

Klíčová slova anglicky

probiotics; quality control; contamination; MOL-PCR; MAGPIX; preamplification; purification

Štítky

14313050, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 19. 1. 2026 12:36, Mgr. Marie Novosadová Šípková, DiS.

Anotace

V originále

Probiotic food supplements are widely consumed, yet their quality and labeling accuracy remain inconsistent due to variable manufacturing and insufficient regulation. Conventional methods, e.g., culture and qPCR, enable detection of only a few microorganisms per assay, restricting comprehensive analysis. To overcome this, we developed a multiplex multiple oligonucleotide ligation PCR (MOL-PCR) assay for simultaneous detection of 29 bacterial and yeast species found in probiotic supplements. Protocol improvements─preamplification, optimized moligo concentrations, and a novel ligation product purification─enhanced sensitivity and reduced background noise. The assay achieved 100% analytical specificity with a detection limit of ∼101–102 microorganisms per reaction. Validation on 35 commercial supplements showed strong concordance with qPCR, with a diagnostic sensitivity of 96.4% and a specificity of 98.1%. Importantly, the method revealed frequent discrepancies between declared and actual product composition, including missing strains and undeclared contaminants. This high-throughput assay provides a robust tool for probiotic quality control and regulatory compliance.

Citovat

KRÁLÍK, Petr; Martin KLANICA a Radka DZIEDZINSKÁ. Development of a Novel Multiplex Multiple Oligonucleotide Ligation PCR System for Identification of Probiotic Bacteria in Food Supplements. JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY. WASHINGTON: AMER CHEMICAL SOC, 2025, roč. 73, č. 51, s. 32901-32912. ISSN 0021-8561. Dostupné z: https://doi.org/10.1021/acs.jafc.5c12300.

@article{2548351,
   author = {Králík, Petr and Klanica, Martin and Dziedzinská, Radka},
   article_location = {WASHINGTON},
   article_number = {51},
   doi = {https://doi.org/10.1021/acs.jafc.5c12300},
   keywords = {probiotics; quality control; contamination; MOL-PCR; MAGPIX; preamplification; purification},
   language = {eng},
   issn = {0021-8561},
   journal = {JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY},
   title = {Development of a Novel Multiplex Multiple Oligonucleotide Ligation PCR System for Identification of Probiotic Bacteria in Food Supplements},
   url = {https://pubs.acs.org/doi/10.1021/acs.jafc.5c12300},
   volume = {73},
   year = {2025}
}

TY  - JOUR
ID  - 2548351
AU  - Králík, Petr - Klanica, Martin - Dziedzinská, Radka
PY  - 2025
TI  - Development of a Novel Multiplex Multiple Oligonucleotide Ligation PCR System for Identification of Probiotic Bacteria in Food Supplements
JF  - JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
VL  - 73
IS  - 51
SP  - 32901-32912
EP  - 32901-32912
PB  - AMER CHEMICAL SOC
SN  - 00218561
KW  - probiotics
KW  - quality control
KW  - contamination
KW  - MOL-PCR
KW  - MAGPIX
KW  - preamplification
KW  - purification
UR  - https://pubs.acs.org/doi/10.1021/acs.jafc.5c12300
N2  - Probiotic food supplements are widely consumed, yet their quality and labeling accuracy remain inconsistent due to variable manufacturing and insufficient regulation. Conventional methods, e.g., culture and qPCR, enable detection of only a few microorganisms per assay, restricting comprehensive analysis. To overcome this, we developed a multiplex multiple oligonucleotide ligation PCR (MOL-PCR) assay for simultaneous detection of 29 bacterial and yeast species found in probiotic supplements. Protocol improvements─preamplification, optimized moligo concentrations, and a novel ligation product purification─enhanced sensitivity and reduced background noise. The assay achieved 100% analytical specificity with a detection limit of ∼101–102 microorganisms per reaction. Validation on 35 commercial supplements showed strong concordance with qPCR, with a diagnostic sensitivity of 96.4% and a specificity of 98.1%. Importantly, the method revealed frequent discrepancies between declared and actual product composition, including missing strains and undeclared contaminants. This high-throughput assay provides a robust tool for probiotic quality control and regulatory compliance.
ER  -

KRÁLÍK, Petr; Martin KLANICA a Radka DZIEDZINSKÁ. Development of a Novel Multiplex Multiple Oligonucleotide Ligation PCR System for Identification of Probiotic Bacteria in Food Supplements. \textit{JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY}. WASHINGTON: AMER CHEMICAL SOC, 2025, roč.~73, č.~51, s.~32901-32912. ISSN~0021-8561. Dostupné z: https://doi.org/10.1021/acs.jafc.5c12300.

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