Development and Inter-Laboratory Validation of Diagnostics
Panel for Detection of Biothreat Bacteria Based on MOL-PCR
Assay

J 2021

Development and Inter-Laboratory Validation of Diagnostics Panel for Detection of Biothreat Bacteria Based on MOL-PCR Assay

JELINKOVA, Pavlina; Jakub HRDÝ; Jirina MARKOVA; Jiri DRESLER; Petr PAJER et al.

Základní údaje

Originální název

Development and Inter-Laboratory Validation of Diagnostics Panel for Detection of Biothreat Bacteria Based on MOL-PCR Assay

Autoři

JELINKOVA, Pavlina; Jakub HRDÝ; Jirina MARKOVA; Jiri DRESLER; Petr PAJER; Oto PAVLIS; Pavel BRANICH; Gabriela BORILOVA; Marketa REICHELOVA; Vladimir BABAK; Nikol RESLOVÁ a Petr KRALIK

Vydání

Microorganisms, Basel, MDPI, 2021, 2076-2607

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10606 Microbiology

Stát vydavatele

Švýcarsko

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 4.926

Označené pro přenos do RIV

Ano

Kód RIV

RIV/00216224:14310/21:00120920

Organizační jednotka

Přírodovědecká fakulta

Klíčová slova anglicky

MOL-PCR; biothreat bacteria; magnetic bead; bioterrorism; detection panel

Štítky

14314010, 14314020, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 16. 2. 2021 11:15, Mgr. Marie Novosadová Šípková, DiS.

Anotace

V originále

Early detection of biohazardous bacteria that can be misused as biological weapons is one of the most important measures to prevent the spread and outbreak of biological warfare. For this reason, many instrument platforms need to be introduced into operation in the field of biological warfare detection. Therefore the purpose of this study is to establish a new detection panel for biothreat bacteria (Bacillus anthracis, Yersinia pestis, Francisella tularensis, and Brucella spp.) and confirm it by collaborative validation by using a multiplex oligonucleotide ligation followed by polymerase chain reaction and hybridization to microspheres by MagPix detection platform (MOL-PCR). Appropriate specific sequences in bacterial DNA were selected and tested to assemble the detection panel, and MOLigo probes (short specific oligonucleotides) were designed to show no cross-reactivity when tested between bacteria and to decrease the background signal measurement on the MagPix platform. During testing, sensitivity was assessed for all target bacteria using serially diluted DNA and was determined to be at least 0.5 ng/mu L. For use as a diagnostic kit and easier handling, the storage stability of ligation premixes (MOLigo probe mixes) was tested. This highly multiplex method can be used for rapid screening to prevent outbreaks arising from the use of bacterial strains for bioterrorism, because time of analysis take under 4 h.

Citovat

JELINKOVA, Pavlina; Jakub HRDÝ; Jirina MARKOVA; Jiri DRESLER; Petr PAJER; Oto PAVLIS; Pavel BRANICH; Gabriela BORILOVA; Marketa REICHELOVA; Vladimir BABAK; Nikol RESLOVÁ a Petr KRALIK. Development and Inter-Laboratory Validation of Diagnostics Panel for Detection of Biothreat Bacteria Based on MOL-PCR Assay. Microorganisms. Basel: MDPI, 2021, roč. 9, č. 1, s. 1-15. ISSN 2076-2607. Dostupné z: https://doi.org/10.3390/microorganisms9010038.

@article{1721199,
   author = {Jelinkova, Pavlina and Hrdý, Jakub and Markova, Jirina and Dresler, Jiri and Pajer, Petr and Pavlis, Oto and Branich, Pavel and Borilova, Gabriela and Reichelova, Marketa and Babak, Vladimir and Reslová, Nikol and Kralik, Petr},
   article_location = {Basel},
   article_number = {1},
   doi = {https://doi.org/10.3390/microorganisms9010038},
   keywords = {MOL-PCR; biothreat bacteria; magnetic bead; bioterrorism; detection panel},
   language = {eng},
   issn = {2076-2607},
   journal = {Microorganisms},
   title = {Development and Inter-Laboratory Validation of Diagnostics Panel for Detection of Biothreat Bacteria Based on MOL-PCR Assay},
   url = {https://doi.org/10.3390/microorganisms9010038},
   volume = {9},
   year = {2021}
}

TY  - JOUR
ID  - 1721199
AU  - Jelinkova, Pavlina - Hrdý, Jakub - Markova, Jirina - Dresler, Jiri - Pajer, Petr - Pavlis, Oto - Branich, Pavel - Borilova, Gabriela - Reichelova, Marketa - Babak, Vladimir - Reslová, Nikol - Kralik, Petr
PY  - 2021
TI  - Development and Inter-Laboratory Validation of Diagnostics Panel for Detection of Biothreat Bacteria Based on MOL-PCR Assay
JF  - Microorganisms
VL  - 9
IS  - 1
SP  - 1-15
EP  - 1-15
PB  - MDPI
SN  - 20762607
KW  - MOL-PCR
KW  - biothreat bacteria
KW  - magnetic bead
KW  - bioterrorism
KW  - detection panel
UR  - https://doi.org/10.3390/microorganisms9010038
L2  - https://doi.org/10.3390/microorganisms9010038
N2  - Early detection of biohazardous bacteria that can be misused as biological weapons is one of the most important measures to prevent the spread and outbreak of biological warfare. For this reason, many instrument platforms need to be introduced into operation in the field of biological warfare detection. Therefore the purpose of this study is to establish a new detection panel for biothreat bacteria (Bacillus anthracis, Yersinia pestis, Francisella tularensis, and Brucella spp.) and confirm it by collaborative validation by using a multiplex oligonucleotide ligation followed by polymerase chain reaction and hybridization to microspheres by MagPix detection platform (MOL-PCR). Appropriate specific sequences in bacterial DNA were selected and tested to assemble the detection panel, and MOLigo probes (short specific oligonucleotides) were designed to show no cross-reactivity when tested between bacteria and to decrease the background signal measurement on the MagPix platform. During testing, sensitivity was assessed for all target bacteria using serially diluted DNA and was determined to be at least 0.5 ng/mu L. For use as a diagnostic kit and easier handling, the storage stability of ligation premixes (MOLigo probe mixes) was tested. This highly multiplex method can be used for rapid screening to prevent outbreaks arising from the use of bacterial strains for bioterrorism, because time of analysis take under 4 h.
ER  -

JELINKOVA, Pavlina; Jakub HRDÝ; Jirina MARKOVA; Jiri DRESLER; Petr PAJER; Oto PAVLIS; Pavel BRANICH; Gabriela BORILOVA; Marketa REICHELOVA; Vladimir BABAK; Nikol RESLOVÁ a Petr KRALIK. Development and Inter-Laboratory Validation of Diagnostics Panel for Detection of Biothreat Bacteria Based on MOL-PCR Assay. \textit{Microorganisms}. Basel: MDPI, 2021, roč.~9, č.~1, s.~1-15. ISSN~2076-2607. Dostupné z: https://doi.org/10.3390/microorganisms9010038.

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