The University of Southampton
University of Southampton Institutional Repository

Amplification refractory mutation system, a highly sensitive and simple polymerase chain reaction assay, for the detection of JAK2 V617F mutation in chronic myeloproliferative disorders

Amplification refractory mutation system, a highly sensitive and simple polymerase chain reaction assay, for the detection of JAK2 V617F mutation in chronic myeloproliferative disorders
Amplification refractory mutation system, a highly sensitive and simple polymerase chain reaction assay, for the detection of JAK2 V617F mutation in chronic myeloproliferative disorders
An acquired mutation in Janus kinase 2 (JAK2), V617F, has recently been identified in human myeloproliferative disorders. Detection of the mutation is helpful in differential diagnosis, prognosis, and predication of therapeutic response. Because the mutation can be present in a small proportion of granulocytic populations in myeloproliferative disorder patients, a highly sensitive detection method is required. In this study, we systematically optimized the reaction conditions of a published amplification refractory mutation system-polymerase chain reaction research protocol to make it a robust clinical diagnostic test. The modifications led to a clear demonstration of the V617F mutation in a patient who would have been easily missed by the original amplification refractory mutation system-polymerase chain reaction assay. The test detects the V617F mutation not only with a high analytic sensitivity of 0.05 to 0.1% but also with a high diagnostic specificity of 99%. In addition, the assay has the ability to distinguish cases with only mutant alleles from cases with mixed normal and mutant alleles. The assay is fast and easy to perform, and no special equipment other than thermocyclers is required. All these features make the assay readily and broadly applicable in clinical molecular diagnostic laboratories.
population, sensitivity and specificity, chronic disease, janus kinase 2, methods, human, patients, polymerase chain reaction, dna mutational analysis, pathology, genetics, alleles, genetic screening, mutation, enzymology, phenylalanine, analysis, research, humans, myeloproliferative disorders, exons, prognosis, laboratories, valine, reproducibility of results, genome, diagnosis
1525-1578
272-276
Chen, Qiaofang
a06c4655-781d-4465-968b-d03617fc1a1b
Lu, Pin
83c3d9ff-6bee-463d-a6fd-6b324e93a398
Jones, Amy V.
3d296088-a099-45cf-b227-541ff59d800c
Cross, Nicholas C.P.
f87650da-b908-4a34-b31b-d62c5f186fe4
Silver, Richard T.
661e0b91-4a5c-4100-9a7a-e4f1db3f3a1d
Wang, Y. Lynn
281a4bfb-c4f7-4bb1-b883-902d17f0e04c
Chen, Qiaofang
a06c4655-781d-4465-968b-d03617fc1a1b
Lu, Pin
83c3d9ff-6bee-463d-a6fd-6b324e93a398
Jones, Amy V.
3d296088-a099-45cf-b227-541ff59d800c
Cross, Nicholas C.P.
f87650da-b908-4a34-b31b-d62c5f186fe4
Silver, Richard T.
661e0b91-4a5c-4100-9a7a-e4f1db3f3a1d
Wang, Y. Lynn
281a4bfb-c4f7-4bb1-b883-902d17f0e04c

Chen, Qiaofang, Lu, Pin, Jones, Amy V., Cross, Nicholas C.P., Silver, Richard T. and Wang, Y. Lynn (2007) Amplification refractory mutation system, a highly sensitive and simple polymerase chain reaction assay, for the detection of JAK2 V617F mutation in chronic myeloproliferative disorders. Journal of Molecular Diagnostics, 9 (2), 272-276. (doi:10.2353/jmoldx.2007.060133).

Record type: Article

Abstract

An acquired mutation in Janus kinase 2 (JAK2), V617F, has recently been identified in human myeloproliferative disorders. Detection of the mutation is helpful in differential diagnosis, prognosis, and predication of therapeutic response. Because the mutation can be present in a small proportion of granulocytic populations in myeloproliferative disorder patients, a highly sensitive detection method is required. In this study, we systematically optimized the reaction conditions of a published amplification refractory mutation system-polymerase chain reaction research protocol to make it a robust clinical diagnostic test. The modifications led to a clear demonstration of the V617F mutation in a patient who would have been easily missed by the original amplification refractory mutation system-polymerase chain reaction assay. The test detects the V617F mutation not only with a high analytic sensitivity of 0.05 to 0.1% but also with a high diagnostic specificity of 99%. In addition, the assay has the ability to distinguish cases with only mutant alleles from cases with mixed normal and mutant alleles. The assay is fast and easy to perform, and no special equipment other than thermocyclers is required. All these features make the assay readily and broadly applicable in clinical molecular diagnostic laboratories.

Full text not available from this repository.

More information

Published date: 1 April 2007
Keywords: population, sensitivity and specificity, chronic disease, janus kinase 2, methods, human, patients, polymerase chain reaction, dna mutational analysis, pathology, genetics, alleles, genetic screening, mutation, enzymology, phenylalanine, analysis, research, humans, myeloproliferative disorders, exons, prognosis, laboratories, valine, reproducibility of results, genome, diagnosis

Identifiers

Local EPrints ID: 59569
URI: https://eprints.soton.ac.uk/id/eprint/59569
ISSN: 1525-1578
PURE UUID: 4f3835d5-0b9e-48a7-80b1-8dec35120c9a
ORCID for Nicholas C.P. Cross: ORCID iD orcid.org/0000-0001-5481-2555

Catalogue record

Date deposited: 04 Sep 2008
Last modified: 20 Jul 2019 01:04

Export record

Altmetrics

Download statistics

Downloads from ePrints over the past year. Other digital versions may also be available to download e.g. from the publisher's website.

View more statistics

Atom RSS 1.0 RSS 2.0

Contact ePrints Soton: eprints@soton.ac.uk

ePrints Soton supports OAI 2.0 with a base URL of https://eprints.soton.ac.uk/cgi/oai2

This repository has been built using EPrints software, developed at the University of Southampton, but available to everyone to use.

We use cookies to ensure that we give you the best experience on our website. If you continue without changing your settings, we will assume that you are happy to receive cookies on the University of Southampton website.

×