The scope of quantitative polymerase chain reaction assays in clinical molecular pathology

Malcomson, Roger D. G.; McCullough, Christian T.; Bruce, David J. Murray; Harrison, DJ

doi:10.1136/mp.48.4.m178

Cited by 5 publications

(4 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Cooperative Oncology clinical trial group used the RT-PCR technique to detect presence of residual cancer cells in CML patients that had undergone therapy (Lee et al, 1989). By the mid-1990s, the analysis of translocation breakpoints entered the era of optimization and quantification (Cross et al, 1994;Malcomson et al, 1995) and techniques developed in one domain were quickly transferred to the analysis of other domains (Lynas et al, 1995 …”

Section: Growth Of the Number Of Papers Indexed In Pubmed With The Tementioning

confidence: 99%

Signs, markers, profiles, and signatures: clinical haematology meets the new genetics (1980–2000)

Keating

Cambrosio

2004

New Genetics and Society

View full text Add to dashboard Cite

Section: Growth Of the Number Of Papers Indexed In Pubmed With The Tementioning

confidence: 99%

Signs, markers, profiles, and signatures: clinical haematology meets the new genetics (1980–2000)

Keating

Cambrosio

2004

New Genetics and Society

View full text Add to dashboard Cite

“…Therefore, to improve DNA recovery and maximise PCR sensitivity, samples here were purified using a commercial silica-based column method [ 61 , 66 ]. For optimal quantitation results, an earlier study showed that DNA should be subjected to PCR within one to two weeks post-extraction [ 67 ]. Here, delay between extraction and testing could have contributed to low DNA loads and positivity ratios in clinical samples.…”

Section: Discussionmentioning

confidence: 99%

Measurement of Epstein-Barr virus DNA load using a novel quantification standard containing two EBV DNA targets and SYBR Green I dye

Lay

Lucas

Ratnamohan

et al. 2010

Virol J

View full text Add to dashboard Cite

BackgroundReactivation of Epstein-Barr virus (EBV) infection may cause serious, life-threatening complications in immunocompromised individuals. EBV DNA is often detected in EBV-associated disease states, with viral load believed to be a reflection of virus activity. Two separate real-time quantitative polymerase chain reaction (QPCR) assays using SYBR Green I dye and a single quantification standard containing two EBV genes, Epstein-Barr nuclear antigen-1 (EBNA-1) and BamHI fragment H rightward open reading frame-1 (BHRF-1), were developed to detect and measure absolute EBV DNA load in patients with various EBV-associated diseases. EBV DNA loads and viral capsid antigen (VCA) IgG antibody titres were also quantified on a population sample.ResultsEBV DNA was measurable in ethylenediaminetetraacetic acid (EDTA) whole blood, peripheral blood mononuclear cells (PBMCs), plasma and cerebrospinal fluid (CSF) samples. EBV DNA loads were detectable from 8.0 × 102 to 1.3 × 108 copies/ml in post-transplant lymphoproliferative disease (n = 5), 1.5 × 103 to 2.0 × 105 copies/ml in infectious mononucleosis (n = 7), 7.5 × 104 to 1.1 × 105 copies/ml in EBV-associated haemophagocytic syndrome (n = 1), 2.0 × 102 to 5.6 × 103 copies/ml in HIV-infected patients (n = 12), and 2.0 × 102 to 9.1 × 104 copies/ml in the population sample (n = 218). EBNA-1 and BHRF-1 DNA were detected in 11.0% and 21.6% of the population sample respectively. There was a modest correlation between VCA IgG antibody titre and BHRF-1 DNA load (rho = 0.13, p = 0.05) but not EBNA-1 DNA load (rho = 0.11, p = 0.11).ConclusionTwo sensitive and specific real-time PCR assays using SYBR Green I dye and a single quantification standard containing two EBV DNA targets, were developed for the detection and measurement of EBV DNA load in a variety of clinical samples. These assays have application in the investigation of EBV-related illnesses in immunocompromised individuals.

show abstract

“…The molecular search for IG and T C R rearrangements may occasionally be the final step in the differential diagnosis between a malignant lymphoma and a nonlymphoid malignancy, either a poorly differentiated neoplasia or, e. g., a neoplasia of "accessory" cells of the immune system (interdigitating and reticulum cell sarcomas) (Raljkiaer et al 1990;Miettinen et al 1993). Far more common in hematologic practice is the valuable confirmation of clonality in lymphoid proliferations with mature Tcell phenotypes, such as cutaneous lymphoid infiltrates and peripheral T-cell lymphomas (Menke et al 1995;Zelickson et al 1991), or the detection of a clonal population admixed with a prominent reactive population, as occurs in the so-called T-cell-rich B-cell lymphoma (Macon et al 1992; Krishnan et al 1994). The identification of a reactive condition underlying an atypical lymphoid population may also rely on the absence of monoclonal IG and TCR rearrangements.…”

mentioning

confidence: 99%

“…Several strategies have been developed in order to establish quantitative PCR methods, the best approach appearing to be the use of internal controls in a competitive PCR. In particular, the use of an identifiable construct that is amplifiable by the same primers as the target under investigation to avoid problems of primer interactions has been successful (Cross 1995;Malcomson et al 1995;Siebert & Larrick 1992). The more frequently used approaches rely on serial dilutions of the PCR products or analyses of multiple replicates, but the nonlinear relationship between the initial target concentration and the resulting concentration of amplicons has to be taken into consideration in order to obtain reliable quantifications (Ouspen-skaia et al 1995).…”

mentioning

confidence: 99%

Molecular pathology in the diagnosis of hematologic neoplasia

Sambade

Sällström

Sundström

1997

APMIS

View full text Add to dashboard Cite

Over the past decade molecular genetic methods have played an increasingly important role in the diagnosis of hematologic malignancies. Moreover, they have provided a tool to analyze many of the non‐random cytogenetic anomalies associated with hematologic neoplasias, contributing considerally to our understanding of several of those diseases, and to improving diagnostic accuracy. The rapid development of molecular genetics progressively allows the replacement of time‐consuming and technically demanding procedures. Even more relevant are the new clinical applications that already include the search for valuable prognostic information and ways of evaluating minimal residual disease or recognizing early relapsing disease. This paper is a critical but necessarily simplified overview of the main contributions of molecular genetics to the field of hematopathology. We discuss the information provided by several molecular methods within different clinical contexts, covering common problems in diagnostic pathology as well as prognostic evaluation and therapy monitoring.

show abstract

The scope of quantitative polymerase chain reaction assays in clinical molecular pathology

Cited by 5 publications

References 37 publications

Signs, markers, profiles, and signatures: clinical haematology meets the new genetics (1980–2000)

Signs, markers, profiles, and signatures: clinical haematology meets the new genetics (1980–2000)

Measurement of Epstein-Barr virus DNA load using a novel quantification standard containing two EBV DNA targets and SYBR Green I dye

Molecular pathology in the diagnosis of hematologic neoplasia

Contact Info

Product

Resources

About