xMAP technology - detection of pathogenic viruses

a 2018

xMAP technology - detection of pathogenic viruses

HRDÝ, Jakub; Petra VAŠÍČKOVÁ; Nikol RESLOVÁ; Veronika HUVAROVÁ; Petr KRÁLÍK et al.

Základní údaje

Originální název

xMAP technology - detection of pathogenic viruses

Autoři

HRDÝ, Jakub; Petra VAŠÍČKOVÁ; Nikol RESLOVÁ; Veronika HUVAROVÁ a Petr KRÁLÍK

Vydání

XXVII. Tomáškovy dny mladých mikrobiologů, 2018

Další údaje

Jazyk

angličtina

Typ výsledku

Konferenční abstrakt

Obor

10606 Microbiology

Stát vydavatele

Česká republika

Utajení

není předmětem státního či obchodního tajemství

Označené pro přenos do RIV

Ano

Kód RIV

RIV/00216224:14310/18:00102994

Organizační jednotka

Přírodovědecká fakulta

ISBN

978-80-210-8963-1

Klíčová slova anglicky

xMAP virus detection multiplex

Příznaky

Mezinárodní význam

Změněno: 31. 1. 2019 10:37, Mgr. Jakub Hrdý, Ph.D.

Anotace

V originále

Most of the currently used reliable methods for molecular detection of microbiological agents rely on systems based on real-time polymerase chain reaction (qPCR). This approach offers sufficiently fast, sensitive and specific way to detect potentially present agents in various samples. Nevertheless, the huge disadvantage of qPCR is limitation of its applications for multiplex formats. There is not enough fluorescent dyes to create truly robust systems for detection of larger number of targets. Based on this disadvantage we utilized the xMAP technology and its open architecture to create high-throughput multiplex systems which could overcome this serious handicap of qPCR. The first step of analysis is ligation. In the presence of target DNA, two specific DNA probes are linked. These probes contain universal primer sequences and one of them also the specific TAG sequence. Next step is monoplex end-point polymerase chain reaction (PCR) allowing the amplification of ligated probes. One of the two used PCR primers is labelled by fluorescent dye. After the amplification, the detection using magnetic beads can follow. These beads are covered with specific oligonucleotides (anti-TAG) and each set of beads has unique absorption spectrum. Amplified probes are thus bound by their specific TAG sequence to complementary anti-TAG sequence on appropriate beads. Beads with bounded probes are eventually sorted using MAGPIX® instrument (Luminex Corporation). By this system it is theoretically possible to detect up to 50 different targets in one reaction. To meet the ever-increasing needs for rapid and reliable detection of viruses in human, food or environmental samples in our laboratory, the multiplex viral panel based on described technology will be developed. Up to now, this system includes probes for detection of hepatitis A, hepatitis E and norovirus. Validation of this system and addition of other possible targets is currently underway.

Citovat

HRDÝ, Jakub; Petra VAŠÍČKOVÁ; Nikol RESLOVÁ; Veronika HUVAROVÁ a Petr KRÁLÍK. xMAP technology - detection of pathogenic viruses. In XXVII. Tomáškovy dny mladých mikrobiologů. 2018. ISBN 978-80-210-8963-1.

@proceedings{1420601,
   author = {Hrdý, Jakub and Vašíčková, Petra and Reslová, Nikol and Huvarová, Veronika and Králík, Petr},
   booktitle = {XXVII. Tomáškovy dny mladých mikrobiologů},
   keywords = {xMAP virus detection multiplex},
   language = {eng},
   isbn = {978-80-210-8963-1},
   title = {xMAP technology - detection of pathogenic viruses},
   year = {2018}
}

TY  - CONF
ID  - 1420601
AU  - Hrdý, Jakub - Vašíčková, Petra - Reslová, Nikol - Huvarová, Veronika - Králík, Petr
PY  - 2018
TI  - xMAP technology - detection of pathogenic viruses
SN  - 9788021089631
KW  - xMAP virus detection multiplex
N2  - Most of the currently used reliable methods for molecular detection of microbiological agents rely on systems based on real-time polymerase chain reaction (qPCR). This approach offers sufficiently fast, sensitive and specific way to detect potentially present agents in various samples. Nevertheless, the huge disadvantage of qPCR is limitation of its applications for multiplex formats. There is not enough fluorescent dyes to create truly robust systems for detection of larger number of targets. Based on this disadvantage we utilized the xMAP technology and its open architecture to create high-throughput multiplex systems which could overcome this serious handicap of qPCR. The first step of analysis is ligation. In the presence of target DNA, two specific DNA probes are linked. These probes contain universal primer sequences and one of them also the specific TAG sequence. Next step is monoplex end-point polymerase chain reaction (PCR) allowing the amplification of ligated probes. One of the two used PCR primers is labelled by fluorescent dye. After the amplification, the detection using magnetic beads can follow. These beads are covered with specific oligonucleotides (anti-TAG) and each set of beads has unique absorption spectrum. Amplified probes are thus bound by their specific TAG sequence to complementary anti-TAG sequence on appropriate beads. Beads with bounded probes are eventually sorted using MAGPIX® instrument (Luminex Corporation). By this system it is theoretically possible to detect up to 50 different targets in one reaction. To meet the ever-increasing needs for rapid and reliable detection of viruses in human, food or environmental samples in our laboratory, the multiplex viral panel based on described technology will be developed. Up to now, this system includes probes for detection of hepatitis A, hepatitis E and norovirus. Validation of this system and addition of other possible targets is currently underway.
ER  -

HRDÝ, Jakub; Petra VAŠÍČKOVÁ; Nikol RESLOVÁ; Veronika HUVAROVÁ a Petr KRÁLÍK. xMAP technology - detection of pathogenic viruses. In \textit{XXVII. Tomáškovy dny mladých mikrobiologů}. 2018. ISBN~978-80-210-8963-1.

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