Evaluation of the Roche LightCycler

Cooper, Photographer: Bob; Cooper, Susan M.; Smith, Julie M; Kitchen, S.; Makris, Michael

doi:10.1097/00001721-200307000-00012

Cited by 13 publications

(1 citation statement)

References 17 publications

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“…In this assay, a post-PCR melting curve analysis is performed using a hybrid probe composed of two sequence-specific probes, including a donor and acceptor. 74 The donor (or sensor) probe is labeled with fluorescein at 3′-end covering the 1,691 position at the F5 gene which is a full match with wild-type allele. The acceptor (anchor or reporter) probe anneals close to the donor probe and is labeled at the 5′-end with LightCycler (LC) Red fluorophore.…”

Section: Dna-based Confirmatory Methodsmentioning

confidence: 99%

Laboratory Diagnosis of Activated Protein C Resistance and Factor V Leiden

Bahraini

Fazeli

Dorgalaleh

2023

Semin Thromb Hemost

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The factor V Leiden (FVL) polymorphism is known as the most common inherited risk factor for venous thrombosis. In turn, FVL is the leading cause of an activated protein C resistance (APCR) phenotype, in which the addition of exogenous activated protein C to plasma does not result in the expected anticoagulant effect. In the routine laboratory approach to the formal diagnosis of FVL, an initial positive screening plasma-based method for APCR is often performed, and only if needed, this is followed by a confirmatory DNA-based assay for FVL. Multiple methods with accepted sensitivity and specificity for determining an APCR/FVL phenotype are commonly categorized into two separate groups: (1) screening plasma-based assays, including qualitative functional clot-based assays, for APCR, and (2) confirmatory DNA-based molecular assays, entailing several tests and platforms, including polymerase chain reaction-based and non-PCR-based techniques, for FVL. This review will describe the methodological aspects of each laboratory test and prepare suggestions on the indication of APCR and FVL testing and method selection.

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Section: Dna-based Confirmatory Methodsmentioning

confidence: 99%

Laboratory Diagnosis of Activated Protein C Resistance and Factor V Leiden

Bahraini

Fazeli

Dorgalaleh

2023

Semin Thromb Hemost

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Analytic validity of genetic tests to identify factor V Leiden and prothrombin G20210A

et al. 2010

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The objective of this study is to systematically review methods for detecting Factor V Leiden or prothrombin G20210A. English-language literature from MEDLINE 1 , EMBASE 1 , The Cochrane Library, the Cumulative Index to Nursing and Allied Health Literature, PsycInfo , 2000-December 2008. Studies assessed methods for detection of these mutations in at least 10 human blood samples and reported concordance, discordance, or reproducibility. Two investigators abstracted data on the sample selection criteria, test operators, DNA extraction, experimental test, reference standard, commercial instruments, concordance rates, explanation of any discordance, and whether discordance resolved after repetition. We assessed strength of the evidence using the GRADE criteria. We reviewed 7,777 titles and included 66 articles. The majority of the reviewed studies used PCR-RFLP or AS-PCR as the reference standard. The studies demonstrated that commercially available and precommercial tests have high analytic validity with all having greater than 99% concordance with the reference standard. With a few exceptions, discordance resolved with repetition of the test, suggesting operator or administrative errors were responsible for the discordant results. In the quality assurance studies, greater than 98% of laboratories demonstrated high, even perfect, accuracy when asked to diagnose a sample with a known mutation. The majority of errors came from a limited number of laboratories. Although not all methods may be accurate, there is high-grade evidence that genetic tests for the detection of FVL and prothrombin G20210A have excellent analytic validity. There is high-grade evidence that most, but not all, clinical laboratories test for FVL and prothrombin G20210A accurately. Am. J. Hematol. 85:264-270, 2010. V

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