Journal of Mass Spectrometry and Advances in the Clinical Lab最新文献

{"title":"The regulatory landscape of laboratory developed tests: Past, present, and a perspective on the future","authors":"Melissa M. Budelier , Jacqueline A. Hubbard","doi":"10.1016/j.jmsacl.2023.02.008","DOIUrl":"10.1016/j.jmsacl.2023.02.008","url":null,"abstract":"","PeriodicalId":52406,"journal":{"name":"Journal of Mass Spectrometry and Advances in the Clinical Lab","volume":"28 ","pages":"Pages 67-69"},"PeriodicalIF":2.2,"publicationDate":"2023-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/59/f0/main.PMC9985058.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10861157","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A customized mass array panel for BCR::ABL1 tyrosine kinase domain mutation screening in chronic myeloid leukemia","authors":"Nittaya Limsuwanachot ,&nbsp;Budsaba Rerkamnuaychoke ,&nbsp;Pimjai Niparuck ,&nbsp;Roongrudee Singdong ,&nbsp;Adcharee Kongruang ,&nbsp;Piyapha Hirunpatrawong ,&nbsp;Thanaporn Siriyakorn ,&nbsp;Pa-thai Yenchitsomanus ,&nbsp;Teerapong Siriboonpiputtana","doi":"10.1016/j.jmsacl.2023.04.002","DOIUrl":"https://doi.org/10.1016/j.jmsacl.2023.04.002","url":null,"abstract":"<div><h3>Introduction</h3><p>The therapeutic strategy and management of chronic myeloid leukemia (CML) have rapidly improved with the discovery of effective tyrosine kinase inhibitors (TKIs) to target BCR::ABL1 oncoprotein. However, nearly 30% of patients develop TKI resistance due to acquired mutations on the tyrosine kinase domain (TKD) of <em>BCR</em>::<em>ABL1</em>.</p></div><div><h3>Methods</h3><p>We customized a mass array panel initially intended to detect and monitor the mutational burden of hotspot <em>BCR</em>::<em>ABL1</em> TKD mutations accumulated in our database, including key mutations recently recommended by European LeukemiaNet. Additionally, we extended the feasibility of using the assay panel for the molecular classification of myeloproliferative neoplasms (MPNs) by incorporating primer sets specific for analyzing <em>JAK2</em> V617F, <em>MPL</em> 515 K/L, and <em>CALR</em> types 1 and 2.</p></div><div><h3>Results</h3><p>We found that the developed mass array panel was superior for detecting and monitoring clinically significant <em>BCR</em>::<em>ABL1</em> TKD mutations, especially in cases with low mutational burden and harboring compound/polyclonal mutations, compared with direct sequencing. Moreover, our customized mass array panel detected common genetic alterations in MPNs, and the findings were consistent with those of other comparable assays available in our laboratory.</p></div><div><h3>Conclusions</h3><p>Our customized mass array panel was practicably used as a routine robust assay for screening and monitoring <em>BCR</em>::<em>ABL1</em> TKD mutations in patients with CML undergoing TKI treatment and feasible for analyzing common genetic mutations in MPNs.</p></div>","PeriodicalId":52406,"journal":{"name":"Journal of Mass Spectrometry and Advances in the Clinical Lab","volume":"28 ","pages":"Pages 122-132"},"PeriodicalIF":2.2,"publicationDate":"2023-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49741227","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Small molecule biomarker discovery: Proposed workflow for LC-MS-based clinical research projects","authors":"S. Rischke ,&nbsp;L. Hahnefeld ,&nbsp;B. Burla ,&nbsp;F. Behrens ,&nbsp;R. Gurke ,&nbsp;T.J. Garrett","doi":"10.1016/j.jmsacl.2023.02.003","DOIUrl":"10.1016/j.jmsacl.2023.02.003","url":null,"abstract":"<div><p>Mass spectrometry focusing on small endogenous molecules has become an integral part of biomarker discovery in the pursuit of an in-depth understanding of the pathophysiology of various diseases, ultimately enabling the application of personalized medicine. While LC-MS methods allow researchers to gather vast amounts of data from hundreds or thousands of samples, the successful execution of a study as part of clinical research also requires knowledge transfer with clinicians, involvement of data scientists, and interactions with various stakeholders.</p><p>The initial planning phase of a clinical research project involves specifying the scope and design, and engaging relevant experts from different fields. Enrolling subjects and designing trials rely largely on the overall objective of the study and epidemiological considerations, while proper pre-analytical sample handling has immediate implications on the quality of analytical data. Subsequent LC-MS measurements may be conducted in a targeted, semi-targeted, or non-targeted manner, resulting in datasets of varying size and accuracy. Data processing further enhances the quality of data and is a prerequisite for in-silico analysis. Nowadays, the evaluation of such complex datasets relies on a mix of classical statistics and machine learning applications, in combination with other tools, such as pathway analysis and gene set enrichment. Finally, results must be validated before biomarkers can be used as prognostic or diagnostic decision-making tools. Throughout the study, quality control measures should be employed to enhance the reliability of data and increase confidence in the results.</p><p>The aim of this graphical review is to provide an overview of the steps to be taken when conducting an LC-MS-based clinical research project to search for small molecule biomarkers.</p></div>","PeriodicalId":52406,"journal":{"name":"Journal of Mass Spectrometry and Advances in the Clinical Lab","volume":"28 ","pages":"Pages 47-55"},"PeriodicalIF":2.2,"publicationDate":"2023-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9982001/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10836640","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Clinical utility of laboratory developed mass spectrometry assays for steroid hormone testing","authors":"Deborah French","doi":"10.1016/j.jmsacl.2023.01.006","DOIUrl":"10.1016/j.jmsacl.2023.01.006","url":null,"abstract":"","PeriodicalId":52406,"journal":{"name":"Journal of Mass Spectrometry and Advances in the Clinical Lab","volume":"28 ","pages":"Pages 13-19"},"PeriodicalIF":2.2,"publicationDate":"2023-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/e1/4b/main.PMC9900367.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10674744","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Analysis of monoclonal immunoglobulins from bone marrow plasma cells using immunopurification and LC-MS","authors":"David R. Barnidge ,&nbsp;Angela Dispenzieri ,&nbsp;Dragan Jevremovic ,&nbsp;David L. Murray","doi":"10.1016/j.jmsacl.2023.04.001","DOIUrl":"10.1016/j.jmsacl.2023.04.001","url":null,"abstract":"<div><h3>Introduction</h3><p>Clonal plasma cells secrete immunoglobulins, each with the exact same amino acid sequence, that are referred to as monoclonal immunoglobulins. The monoclonal heavy chain and light chain secreted from clonal plasma cells have the same molecular mass prior to the addition of post-translational modifications (PTMs) since their amino acid sequences are the same.</p></div><div><h3>Objective</h3><p>To examine the molecular masses of monoclonal light chains and heavy chains isolated directly from the cytoplasm of bone marrow (BM) plasma cells and compare them to the serum derived monoclonal heavy and light chains.</p></div><div><h3>Methods</h3><p>Using immunopurification and LC-MS we compared the molecular masses of immunoglobulins immunopurified from a patient’s serum to those immunopurified from the cytoplasm of their BM plasma cells.</p></div><div><h3>Results</h3><p>Our findings demonstrate that the light chain molecular masses were identical whether they were obtained from serum or plasma cell cytoplasm. However, the heavy chain molecular masses did not match in bone marrow and serum due to differences in glycosylation, a common post-translational modification (PTM) found on the heavy chain.</p></div><div><h3>Conclusion</h3><p>The data presented here show that by using LC-MS to analyze monoclonal immunoglobulins (also referred to as miRAMM) additional phenotype information is obtained at the cellular level which is complementary to other more common techniques such as flow cytometry and histopathology.</p></div>","PeriodicalId":52406,"journal":{"name":"Journal of Mass Spectrometry and Advances in the Clinical Lab","volume":"28 ","pages":"Pages 133-141"},"PeriodicalIF":2.2,"publicationDate":"2023-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/04/15/main.PMC10149385.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9627712","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Pediatric laboratory developed tests filling the gaps for children in crisis","authors":"Dustin R. Bunch","doi":"10.1016/j.jmsacl.2023.02.012","DOIUrl":"10.1016/j.jmsacl.2023.02.012","url":null,"abstract":"","PeriodicalId":52406,"journal":{"name":"Journal of Mass Spectrometry and Advances in the Clinical Lab","volume":"28 ","pages":"Pages 80-81"},"PeriodicalIF":2.2,"publicationDate":"2023-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/45/0c/main.PMC9993020.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9146980","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A persistently febrile patient post-bone marrow transplant","authors":"Ashley R. Rackow,&nbsp;Claire E. Knezevic","doi":"10.1016/j.jmsacl.2023.01.005","DOIUrl":"10.1016/j.jmsacl.2023.01.005","url":null,"abstract":"","PeriodicalId":52406,"journal":{"name":"Journal of Mass Spectrometry and Advances in the Clinical Lab","volume":"28 ","pages":"Pages 9-12"},"PeriodicalIF":2.2,"publicationDate":"2023-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9925957/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10741982","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Pre-analytical conditions influencing analysis of folate in dried plasma microsamples","authors":"Christopher M. Shuford,&nbsp;Evan W. McConnell,&nbsp;Stacy Dee,&nbsp;Russell P. Grant","doi":"10.1016/j.jmsacl.2023.01.003","DOIUrl":"10.1016/j.jmsacl.2023.01.003","url":null,"abstract":"<div><h3>Introduction</h3><p>Determination of folate insufficiency is of considerable interest given its importance in fetal development and red blood cell formation; however, access to blood tests may be limited due to the requirement for phlebotomy as well as controlled temperature shipping of blood specimens to laboratories for testing due to the inherent instability of folate and its vitamers.</p></div><div><h3>Methods</h3><p>An LC-MS/MS test was developed and validated for the measurement of 5-methyltetrahydrofolate (5MTHF) in dried plasma specimens collected from fingerstick blood using a laminar flow blood separation device, as well as liquid venous plasma for comparison. Two pre-analytical factors investigated influencing the measurement of 5MTHF in dried plasma were hemolysis of the fingerstick blood during collection and storage/shipment of the dried plasma.</p></div><div><h3>Results</h3><p>Although observed infrequently, hemolysis &gt;10 % resulted in elevated 5MTHF measurements, but hemolysis &gt;1 % resulted in elevated chloride measurements, which were necessary to normalize 5MTHF measurements for variation in volume of dried plasma specimens. Stability of 5MTHF was improved in dried plasma relative to liquid plasma at ambient temperatures, but not sufficiently to allow for uncontrolled temperature shipping despite controlling for humidity and light exposure. Shipping studies emulating ISTA procedure 7D were conducted with a reusable cold packaging solution. The packaging failed to stabilize 5MTHF in dried plasma specimens during a 2-day summer shipping evaluation, but did provide sufficient temperature control to stabilize 5MTHF during the overnight shipping evaluation.</p></div><div><h3>Conclusion</h3><p>Our studies provide boundary conditions with respect to hemolysis, storage, and shipping for successful analysis of 5MTHF from dried plasma specimens.</p></div>","PeriodicalId":52406,"journal":{"name":"Journal of Mass Spectrometry and Advances in the Clinical Lab","volume":"28 ","pages":"Pages 1-8"},"PeriodicalIF":2.2,"publicationDate":"2023-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/a6/97/main.PMC9894916.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10652069","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Evaluating the performance of the Roche FEN2 fentanyl immunoassay and its clinical implementation: The role of LDT-based mass spectrometry testing","authors":"Marlen Menlyadiev ,&nbsp;Raymond T. Suhandynata ,&nbsp;Kyle Lund ,&nbsp;Michael J. Kelner ,&nbsp;Robert L. Fitzgerald","doi":"10.1016/j.jmsacl.2023.02.009","DOIUrl":"10.1016/j.jmsacl.2023.02.009","url":null,"abstract":"<div><h3>Introduction</h3><p>While laboratory-developed tests (LDTs) using liquid chromatography tandem mass spectrometry (LC-MS/MS) are widely employed to support the development of FDA-cleared drug immunoassays, their significance in the clinical implementation and evaluation of such assays is often overlooked. This paper reports on the important role of LC-MS/MS LDTs in demonstrating improved performance of the Roche FEN2 fentanyl immunoassay compared with the Thermo DRI fentanyl immunoassay.</p></div><div><h3>Methods</h3><p>The FEN2 assay was implemented according to the manufacturer's instructions and its performance was compared to the existing DRI assay using LC-MS/MS as a reference. Clinical sensitivity and specificity were determined using 250 consecutive random patient specimens. Spiking experiments were conducted to determine cross-reactivity with 31 fentanyl analogs. Select DRI false-positive samples were analyzed by the FEN2 assay via time-of-flight mass spectrometry method (LC-QTOF).</p></div><div><h3>Results</h3><p>The FEN2 assay showed improved clinical sensitivity compared to the DRI (98% vs 61%) in 250 consecutive patient samples due to its ability to detect norfentanyl. It also showed better clinical specificity by correctly classifying select DRI false-positive results. Upon implementation in clinical practice, the FEN2 resulted in a higher screening positivity rate than the DRI (17.3% vs 13.3%) and a greater LC-MS/MS confirmation rate of immunoassay-positive samples (96.8% vs 88.8%, respectively).</p></div><div><h3>Conclusion</h3><p>The use of LC-MS/MS LDTs demonstrated that the FEN2 assay has greater clinical sensitivity and is less prone to false-positives than the DRI assay. These findings support the use of FEN2 in routine clinical practice and emphasize the role of mass spectrometry-based LDTs in clinical toxicology testing.</p></div>","PeriodicalId":52406,"journal":{"name":"Journal of Mass Spectrometry and Advances in the Clinical Lab","volume":"28 ","pages":"Pages 105-113"},"PeriodicalIF":2.2,"publicationDate":"2023-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10070886/pdf/main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9270735","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparison of two highly sensitive benzodiazepine immunoassay lab developed tests for urine drug testing in clinical specimens","authors":"Kyle Lund ,&nbsp;Marlen Menlyadiev ,&nbsp;Kyunghoon Lee ,&nbsp;Michael J. Kelner ,&nbsp;Robert L. Fitzgerald ,&nbsp;Raymond T. Suhandynata","doi":"10.1016/j.jmsacl.2023.02.010","DOIUrl":"10.1016/j.jmsacl.2023.02.010","url":null,"abstract":"<div><h3>Background</h3><p>The VALID Act is a legislative effort that, if enacted, would alter the regulatory requirements of laboratory developed tests (LDTs) used for clinical testing in the United States. Benzodiazepines, which are primarily excreted into urine as glucuronidated metabolites such as lorazepam, cross-react poorly with FDA-cleared immunoassays, leading to false-negatives. This shortfall can be addressed with LDTs created by adding glucuronidase to the immunoassay reagents producing “high sensitivity” assays that detect glucuronidated metabolites.</p></div><div><h3>Methods</h3><p>Precision and stability of two high-sensitivity (HS) benzodiazepine immunoassays from Roche and Thermo Scientific were evaluated using manufacturer-supplied quality control (QC) material and glucuronidated QC material. The immunoassays were directly compared to an LC-MS/MS LDT benzodiazepine assay to determine clinical sensitivity/specificity using urine specimens (n = 82 for Thermo Scientific; n = 265 for Roche). The clinical impact of the HS LDT immunoassay was determined by analyzing clinical testing results 60 days before and after its implementation.</p></div><div><h3>Results</h3><p>The precision and clinical sensitivity/specificity of the HS-Thermo Scientific and HS-Roche benzodiazepine assays were acceptable. The reagent stability of the HS-Thermo Scientific immunoassay was poor, whereas the HS-Roche immunoassay was stable. After implementation of the HS-Roche benzodiazepine immunoassay as an LDT, there was a 30-fold increase <em>(p</em>-value: &lt; 0.00001) in the percentage of lorazepam confirmations.</p></div><div><h3>Conclusions</h3><p>We demonstrate the development and validation of an immunoassay LDT with improved sensitivity for glucuronidated benzodiazepines. This LDT can detect glucuronidated benzodiazepines in clinical urine specimens and is stable for 60 days. Importantly, we were able to validate the immunoassay as an LDT by utilizing an LC-MS/MS LDT.</p></div>","PeriodicalId":52406,"journal":{"name":"Journal of Mass Spectrometry and Advances in the Clinical Lab","volume":"28 ","pages":"Pages 91-98"},"PeriodicalIF":2.2,"publicationDate":"2023-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/14/f3/main.PMC10020650.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9152995","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}