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E. coli from drinking water
Published in Cara Gleeson, Nick Gray, The Coliform Index and Waterborne Disease, 1996
The use of a multiplex PCR for amplification and detection of more than one target organism can be useful for monitoring multiple microbial pathogens in a single water sample (Bej et al., 1991). Multiplex PCR is a procedure whereby two or more primers are used to amplify two or more target sequences, and has been adapted so as to allow the detection of gene sequences associated with different groups of bacteria in environmental samples (Lang et al., 1994), as it is possible that water samples will contain more than one type of microbial pathogen in addition to indicator organisms. A multiplex using lac-Z, lamB, uid and ss rRNA permits the simultaneous detection of a number of micro-organisms including coliforms, E. coli and the enteric pathogens Salmonella, Shigella and Legionella (Bej et al., 1991).
Genes and genomics
Published in Firdos Alam Khan, Biotechnology Fundamentals, 2018
Multiplex PCR consists of multiple primer sets within a single PCR mixture to produce amplicons of varying sizes that are specific to different DNA sequences. By targeting multiple genes at once, additional information may be gained from a single test run that otherwise would require several times the reagents and more time to perform. Annealing temperatures for each of the primer sets must be optimized to work correctly within a single reaction, and amplicon sizes (i.e., their base pair length) should be different enough to form distinct bands when visualized by gel electrophoresis.
Principles and Techniques for Deoxyribonucleic Acid (DNA) Manipulation
Published in Hajiya Mairo Inuwa, Ifeoma Maureen Ezeonu, Charles Oluwaseun Adetunji, Emmanuel Olufemi Ekundayo, Abubakar Gidado, Abdulrazak B. Ibrahim, Benjamin Ewa Ubi, Medical Biotechnology, Biopharmaceutics, Forensic Science and Bioinformatics, 2022
Nwadiuto (Diuto) Esiobu, Ifeoma M. Ezeonu, Francisca Nwaokorie
Multiplex PCR – Multiplex PCR is a variant of PCR, which allows the simultaneous amplification of many DNA targets in one reaction by applying different primer pairs in the same reaction. For instance, it could be used to detect the presence of several virulence genes from the DNA extracted from the pure culture of a pathogen. It is also used in virology to detect different types of viruses from a sample. The different specific primers used in the reaction are able to specifically find their targets and mark them for amplification.
Detection of extended spectrum beta-lactamase genes in strains of Escherichia coli and Klebsiella pneumoniae isolated from recreational water and tertiary hospital waste water in Zaria, Nigeria
Published in International Journal of Environmental Health Research, 2022
H. I. Atta, S. M. Idris, B. H. Gulumbe, O. J. Awoniyi
Colonies from a 24-hour culture of the ESBL-producing isolates were aseptically suspended in nuclease-free microtubes containing 100 µL of nuclease-free water. The bacterial DNA was extracted from the suspension using Qiagen DNAeasy extraction kit (Qiagen, Germany) following manufacturer’s instructions. A multiplex PCR reaction was used in amplifying the following genes: TEM, SHV and CTX-M (Bali et al. 2010).