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10.3390/ijms22116061

http://scihub22266oqcxt.onion/10.3390/ijms22116061
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34199760!8199996!34199760
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suck abstract from ncbi


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pmid34199760      Int+J+Mol+Sci 2021 ; 22 (11): ä
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  • 3 Tth Endonuclease Cleavage Polymerase Chain Reaction (3TEC-PCR) Technology for Single-Base-Specific Multiplex Pathogen Detection using a Two-Oligonucleotide System #MMPMID34199760
  • Higgins O; Smith TJ
  • Int J Mol Sci 2021[Jun]; 22 (11): ä PMID34199760show ga
  • Polymerase chain reaction (PCR) is the standard in nucleic acid amplification technology for infectious disease pathogen detection and has been the primary diagnostic tool employed during the global COVID-19 pandemic. Various PCR technology adaptations, typically using two-oligonucleotide dye-binding methods or three-oligonucleotide hydrolysis probe systems, enable real-time multiplex target detection or single-base specificity for the identification of single-nucleotide polymorphisms (SNPs). A small number of two-oligonucleotide PCR systems facilitating both multiplex detection and SNP identification have been reported; however, these methods often have limitations in terms of target specificity, production of variable or false-positive results, and the requirement for extensive optimisation or post-amplification analysis. This study introduces 3' Tth endonuclease cleavage PCR (3TEC-PCR), a two-oligonucleotide PCR system incorporating a modified primer/probe and a thermostable cleavage enzyme, Tth endonuclease IV, for real-time multiplex detection and SNP identification. Complete analytical specificity, low limits of detection, single-base specificity, and simultaneous multiple target detection have been demonstrated in this study using 3TEC-PCR to identify bacterial meningitis associated pathogens. This is the first report of a two-oligonucleotide, real-time multiplex PCR technology with single-base specificity using Tth endonuclease IV.
  • |*Polymorphism, Single Nucleotide[MESH]
  • |Alleles[MESH]
  • |DNA, Bacterial/genetics/isolation & purification/metabolism[MESH]
  • |DNA-Directed DNA Polymerase/*metabolism[MESH]
  • |Haemophilus influenzae/genetics[MESH]
  • |Humans[MESH]
  • |Meningitis, Bacterial/diagnosis/microbiology[MESH]
  • |Neisseria meningitidis/genetics[MESH]
  • |Polymerase Chain Reaction/*methods[MESH]


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