Polymerase Chain Reaction: Types, Utilities and Limitations

Hernández-Rodríguez, Patricia; Ramírez, Arlen Gómez

doi:10.5772/37450

Cited by 5 publications

(3 citation statements)

References 87 publications

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“…Conventional endpoint assays, in particular nested formats, are mostly used for amplifying products for sequencing. While these nested RT-PCRs are very sensitive and allow amplification also in cases of low viral load and from suboptimal specimens such as serum, they require strict contamination control measures [46]. The same clinical specimens may be used for measles and rubella virus detection, facilitating simultaneous or subsequent investigation.…”

Section: Detection Of Viral Rnamentioning

confidence: 99%

Challenges of measles and rubella laboratory diagnostic in the era of elimination

Hübschen

Bork

Brown

et al. 2017

Clinical Microbiology and Infection

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The Member States of the WHO European Region adopted the goal of measles and rubella elimination more than 10 years ago, but so far only 21 of 53 countries have reached this target. Laboratory investigation of suspected cases is essential to support disease elimination efforts. Therefore, WHO maintains a network of accredited laboratories providing high-quality testing. Laboratory investigation heavily relies on specific IgM serology and increasingly on virus detection by reverse transcription (RT)-PCR, but other methods such as IgG avidity testing and genetic characterization of virus strains have gained in importance. In elimination settings, often few samples from suspected cases are available for testing, but testing proficiency must be maintained. The predictive value of an IgM-positive result decreases and other rash-fever disease aetiologies become more important. In addition, cases with a rash after measles/rubella vaccination or with mild disease after waning of vaccine-induced antibodies are seen more often. Thus, it is necessary to perform comprehensive and potentially time-consuming and costly investigations of every suspected case using quality-controlled laboratory methods. At the same time rapid feedback to public health officers is required for timely interventions. The introduction of new laboratory methods for comprehensive case investigations requires training of staff under the supervision of WHO-accredited reference laboratories and the definition of appropriate test algorithms. Clinical, laboratory, and epidemiological data are essential for final case classification and investigation of chains of transmission in the endgame of measles and rubella elimination.

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Section: Detection Of Viral Rnamentioning

confidence: 99%

Challenges of measles and rubella laboratory diagnostic in the era of elimination

Hübschen

Bork

Brown

et al. 2017

Clinical Microbiology and Infection

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show abstract

“…Besides this, it is used for sample amplification, gene detection and gene‐mapping of many living beings as well as extinct organisms such as animals, plants, viruses, and bacteria based on their genetic similarities. Due to this, many complex organisms could be classified accordingly on their phylogenetic origins and thus, provide excellent insights into the genealogy of species (Schrader et al, ; Hernández‐Rodríguez and Ramirez, ). Currently, PCR is commonly used in clinical settings as a diagnostic tool to identify a range of diseases including different types of cancers and genetic disorders (Wang et al, ; Liu et al, ).…”

Section: Polymerase Chain Reactionmentioning

confidence: 99%

The evolution of molecular diagnosis using digital polymerase chain reaction to detect cancer via cell‐free DNA and circulating tumor cells

Melo-Silva

Lucena

Hueneburg

2019

Cell Biology International

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Cancer is one of the most important causes of death worldwide. The onset of cancer may be initiated due to a variety of factors such as environment, genetics or even due to personal lifestyle choices. To counteract this tremendous increase, the demand for a new technology has risen. By this means, the use of digital polymerase chain reaction (dPCR) has been shown to be a promising methodology in the early detection of many types of cancers. Furthermore, several researchers confirmed that the use of tumor cell‐free DNA (cfDNA) and circulating tumor cells (CTC) in peripheral blood is essential in revealing an early prognosis of such diseases. Besides this, it was established that dPCR might be used in a much more efficient, accurate, and reliable manner to amplify a variety of genetic material up to the identification of mutations in hematological diseases. Therefore, this article demonstrates the differences between conventional PCR and dPCR as a molecular technique to detect the early onset of cancer. Furthermore, CTC and cfDNA were officially approved by the Food and Drug Administration as new biological biomarkers in cancer development and monitoring.

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“…In recent years, modifications or variants have been developed from the basic PCR method to improve performance and specificity, and to achieve the amplification of other molecules of interest in research as RNA [117]. Some of these variants are:…”

Section: Ivc Some Examples Of Pcr Methods Currently Usedmentioning

confidence: 99%

Η Κλινική Εφαρμογή Των Μοριακών Τεχνικών Στη Διάγνωση Της Ορθοπαιδικής Λοίμωξης

Zegaer¹,

Ζιγκαέρ²

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Intraoperative conventional bacteriological cultures were compared with different polymerasechain reaction (PCR) methods in patients with total joint arthroplasties. The isolated bacteria wereinvestigated for biofilm formation, and the biofilm forming strains, in their planktonic and biofilmforms, were further tested for their antimicrobial resistance against several clinically importantantimicrobials. Forty four bone and joint samples were included and classified as infected or noninfectedaccording to standard criteria for periprosthetic hip and knee infections. For thebacteriological diagnosis, conventional culture, two types of universal PCR and species specificPCR for three selected pathogens (S. aureus, S. epidermidis, and P. aeruginosa) were applied.Biofilm formation determination was performed by the tissue culture plate method. Antimicrobialsusceptibility of the planktonic bacteria was performed by the minimal inhibitory concentrationdetermination and, of the biofilm forms, by the minimal inhibitory concentration for bacterial regrowthfrom the biofilm. Twenty samples were culture positive, with S. epidermidis, S. aureus orP. aeruginosa. All PCR methods were very ineffective in detecting only one pathogen. All isolateswere biofilm positive and their biofilm forms were highly resistant. In this study, compared toPCR, culture remains the “gold standard”. The biofilm formation by the causative bacteria and theconcomitant manifold increased antimicrobial resistance may explain the clinical failure oftreatment in some cases and should be considered in the future for therapeutic planning.

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Polymerase Chain Reaction: Types, Utilities and Limitations

Cited by 5 publications

References 87 publications

Challenges of measles and rubella laboratory diagnostic in the era of elimination

Challenges of measles and rubella laboratory diagnostic in the era of elimination

The evolution of molecular diagnosis using digital polymerase chain reaction to detect cancer via cell‐free DNA and circulating tumor cells

Η Κλινική Εφαρμογή Των Μοριακών Τεχνικών Στη Διάγνωση Της Ορθοπαιδικής Λοίμωξης

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