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Assay
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==General steps== An assay (analysis) is never an isolated process, as it must be accompanied with pre- and post-analytic procedures. Both the communication order (the request to perform an assay plus related information) ''and'' the handling of the specimen itself (the collecting, documenting, transporting, and processing done before beginning the assay) are pre-analytic steps. Similarly, after the assay is completed the results must be documented, verified and communicated—the post-analytic steps. As with any multi-step [[information]] handling and [[Information transmission|transmission]] system, the variation and errors in reporting final results entail not only those intrinsic to the assay itself but also those occurring in the pre-analytic and post-analytic procedures. While the analytic steps of the assay itself get much attention,<ref>{{cite journal|last1=Bonini|first1=P|last2=Plebani|first2=M|last3=Ceriotti|first3=F|last4=Rubboli|first4=F|title=Errors in laboratory medicine.|journal=Clinical Chemistry|date=May 2002|volume=48|issue=5|pages=691–8|doi=10.1093/clinchem/48.5.691|pmid=11978595|doi-access=free}}</ref> it is those that get less attention of the chain of users—the pre-analytic and post-analytic procedures—that typically accumulate the most errors; e.g., pre-analytic steps in medical laboratory assays may contribute 32–75% of all lab errors.<ref>{{cite journal|last1=Hammerling|first1=Julie A.|title=A Review of Medical Errors in Laboratory Diagnostics and Where We Are Today: Table 1|journal=Laboratory Medicine|date=1 February 2012|volume=43|issue=2|pages=41–44|doi=10.1309/LM6ER9WJR1IHQAUY|doi-access=free}}</ref> Assays can be very diverse, but generally involve the following general steps: # '''Sample processing and manipulation''' in order to selectively present the target in a discernible or measurable form to a discrimination/identification/detection system. It might involve a simple centrifugal separation or washing or filtration or capture by some form of selective binding or it may even involve modifying the target e.g. epitope retrieval in immunological assays or cutting down the target into pieces e.g. in [[Mass Spectrometry]]. Generally there are multiple separate steps done before an assay and are called preanalytic processing. But some of the manipulations may be inseparable part of the assay itself and will not thus be considered pre-analytic. # '''Target-specific discrimination/identification principle''': to discriminate from background (noise) of similar components and specifically identify a particular target component ("analyte") in a biological material by its specific attributes. (e.g. in a [[Polymerase chain reaction|PCR]] assay a specific oligonucleotide primer identifies the target by [[base pairing]] based on the specific nucleotide sequence unique to the target). # '''Signal (or target) amplification system''': The presence and quantity of that analyte is converted into a detectable signal generally involving some method of signal amplification, so that it can be easily discriminated from noise and measured - e.g. in a [[Polymerase chain reaction|PCR]] assay among a mixture of DNA sequences only the specific target is amplified into millions of copies by a [[DNA polymerase]] enzyme so that it can be discerned as a more prominent component compared to any other potential components. Sometimes the concentration of the analyte is too large and in that case the assay may involve sample dilution or some sort of signal diminution system which is a negative amplification. # '''Signal detection (and interpretation) system''': A system of deciphering the amplified signal into an interpretable output that can be quantitative or qualitative. It can be visual or manual very crude methods or can be very sophisticated electronic digital or analog detectors. #'''Signal enhancement and noise filtering''' may be done at any or all of the steps above. Since the more downstream a step/process during an assay, the higher the chance of carrying over noise from the previous process and amplifying it, multiple steps in a sophisticated assay might involve various means of signal-specific sharpening/enhancement arrangements and noise reduction or filtering arrangements. These may simply be in the form of a narrow [[band-pass]] optical filter, or a blocking reagent in a binding reaction that prevents nonspecific binding or a [[quencher|quenching]] reagent in a fluorescence detection system that prevents "autofluorescence" of background objects. {{citation needed|date=September 2013}}
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