Marcello Viscovo, Mia de Laurent Clemmensen, Federica Fosso, Elena Maiolo, Francesco Autore, Luca Laurenti, Stefan Hohaus, Patrizia Chiusolo
{"title":"PlentiPlex™ MYD88 Waldenström 淋巴瘤 qPCR 分析：检测 MYD88 L265P 突变的高灵敏度方法。","authors":"Marcello Viscovo, Mia de Laurent Clemmensen, Federica Fosso, Elena Maiolo, Francesco Autore, Luca Laurenti, Stefan Hohaus, Patrizia Chiusolo","doi":"10.1111/ijlh.14255","DOIUrl":null,"url":null,"abstract":"<div>\n \n \n <section>\n \n <h3> Introduction</h3>\n \n <p>Agarose gel-based conventional and real-time allele-specific polymerase chain reaction (AS-PCR) assays are currently used for sensitive detection and quantification of <i>MYD88</i> L265P mutation. Visual inspection of an agarose gel can often be ambiguous. We propose a new allele-specific quantification PCR (AS-qPCR) assay, PlentiPlex™ <i>MYD</i>88 Waldenström lymphoma qPCR assay, that uses Intercalating Nucleic Acid (INA®) technology for increased affinity and specificity.</p>\n </section>\n \n <section>\n \n <h3> Methods</h3>\n \n <p>This study compares PlentiPlex™ <i>MYD</i>88 Waldenström lymphoma qPCR assay with conventional AS-PCR. We included a total of 102 peripheral and bone marrow blood samples from 94 patients with a lymphoproliferative disorder. Droplet digital PCR (ddPCR) was used as a third method in case of discrepancy.</p>\n </section>\n \n <section>\n \n <h3> Results</h3>\n \n <p>A positive percent agreement of 100% (95% CI 0.92–1.0) and a negative percent agreement of 98% (95% CI 0.90–1.0) were found between the conventional AS-PCR and the AS-qPCR methods. Including the ddPCR results to validate the discrepant cases, the sensitivity and specificity of PlentiPlex™ <i>MYD</i>88 Waldenström lymphoma qPCR Assay were 1.0 (95% CI 0.97–1.0) and 1.0 (95% CI 0.96–1.0), respectively.</p>\n </section>\n \n <section>\n \n <h3> Conclusion</h3>\n \n <p>Our data demonstrate that PlentiPlex™ <i>MYD</i>88 Waldenström lymphoma qPCR assay is a fast, highly sensitive, and specific method for the detection of <i>MYD</i>88 L265P compared with conventional AS-PCR.</p>\n </section>\n </div>","PeriodicalId":14120,"journal":{"name":"International Journal of Laboratory Hematology","volume":null,"pages":null},"PeriodicalIF":2.2000,"publicationDate":"2024-02-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"PlentiPlex™ MYD88 Waldenström lymphoma qPCR assay: A highly sensitive method for detection of MYD88 L265P mutation\",\"authors\":\"Marcello Viscovo, Mia de Laurent Clemmensen, Federica Fosso, Elena Maiolo, Francesco Autore, Luca Laurenti, Stefan Hohaus, Patrizia Chiusolo\",\"doi\":\"10.1111/ijlh.14255\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div>\\n \\n \\n <section>\\n \\n <h3> Introduction</h3>\\n \\n <p>Agarose gel-based conventional and real-time allele-specific polymerase chain reaction (AS-PCR) assays are currently used for sensitive detection and quantification of <i>MYD88</i> L265P mutation. Visual inspection of an agarose gel can often be ambiguous. We propose a new allele-specific quantification PCR (AS-qPCR) assay, PlentiPlex™ <i>MYD</i>88 Waldenström lymphoma qPCR assay, that uses Intercalating Nucleic Acid (INA®) technology for increased affinity and specificity.</p>\\n </section>\\n \\n <section>\\n \\n <h3> Methods</h3>\\n \\n <p>This study compares PlentiPlex™ <i>MYD</i>88 Waldenström lymphoma qPCR assay with conventional AS-PCR. We included a total of 102 peripheral and bone marrow blood samples from 94 patients with a lymphoproliferative disorder. Droplet digital PCR (ddPCR) was used as a third method in case of discrepancy.</p>\\n </section>\\n \\n <section>\\n \\n <h3> Results</h3>\\n \\n <p>A positive percent agreement of 100% (95% CI 0.92–1.0) and a negative percent agreement of 98% (95% CI 0.90–1.0) were found between the conventional AS-PCR and the AS-qPCR methods. 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PlentiPlex™ MYD88 Waldenström lymphoma qPCR assay: A highly sensitive method for detection of MYD88 L265P mutation
Introduction
Agarose gel-based conventional and real-time allele-specific polymerase chain reaction (AS-PCR) assays are currently used for sensitive detection and quantification of MYD88 L265P mutation. Visual inspection of an agarose gel can often be ambiguous. We propose a new allele-specific quantification PCR (AS-qPCR) assay, PlentiPlex™ MYD88 Waldenström lymphoma qPCR assay, that uses Intercalating Nucleic Acid (INA®) technology for increased affinity and specificity.
Methods
This study compares PlentiPlex™ MYD88 Waldenström lymphoma qPCR assay with conventional AS-PCR. We included a total of 102 peripheral and bone marrow blood samples from 94 patients with a lymphoproliferative disorder. Droplet digital PCR (ddPCR) was used as a third method in case of discrepancy.
Results
A positive percent agreement of 100% (95% CI 0.92–1.0) and a negative percent agreement of 98% (95% CI 0.90–1.0) were found between the conventional AS-PCR and the AS-qPCR methods. Including the ddPCR results to validate the discrepant cases, the sensitivity and specificity of PlentiPlex™ MYD88 Waldenström lymphoma qPCR Assay were 1.0 (95% CI 0.97–1.0) and 1.0 (95% CI 0.96–1.0), respectively.
Conclusion
Our data demonstrate that PlentiPlex™ MYD88 Waldenström lymphoma qPCR assay is a fast, highly sensitive, and specific method for the detection of MYD88 L265P compared with conventional AS-PCR.
期刊介绍：
The International Journal of Laboratory Hematology provides a forum for the communication of new developments, research topics and the practice of laboratory haematology.
The journal publishes invited reviews, full length original articles, and correspondence.
The International Journal of Laboratory Hematology is the official journal of the International Society for Laboratory Hematology, which addresses the following sub-disciplines: cellular analysis, flow cytometry, haemostasis and thrombosis, molecular diagnostics, haematology informatics, haemoglobinopathies, point of care testing, standards and guidelines.
The journal was launched in 2006 as the successor to Clinical and Laboratory Hematology, which was first published in 1979. An active and positive editorial policy ensures that work of a high scientific standard is reported, in order to bridge the gap between practical and academic aspects of laboratory haematology.
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