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  1. Article: Applications of Digital Polymerase Chain Reaction (dPCR) in Molecular and Clinical Testing.

    Wainman, Lauren M / Sathyanarayana, Shivaprasad H / Lefferts, Joel A

    The journal of applied laboratory medicine

    2024  Volume 9, Issue 1, Page(s) 124–137

    Abstract: Background: Digital polymerase chain reaction (dPCR) is an accurate and sensitive molecular method that can be used in clinical diagnostic, prognostic, and predictive tests. The key component of the dPCR method is the partitioning of a single reaction ... ...

    Abstract Background: Digital polymerase chain reaction (dPCR) is an accurate and sensitive molecular method that can be used in clinical diagnostic, prognostic, and predictive tests. The key component of the dPCR method is the partitioning of a single reaction into many thousands of droplets, nanochannels or other nano- or picoliter-sized reactions. This results in high enough sensitivity to detect rare nucleic acid targets and provides an absolute quantification of target sequences or alleles compared to other PCR-based methods.
    Content: An increasing number of dPCR platforms have been introduced commercially in recent years and more are being developed. These platforms differ in the method of partitioning, degree of automation, and multiplexing capabilities but all can be used in similar ways for sensitive and highly accurate quantification of a variety of nucleic acid targets. Currently, clinical applications of dPCR include oncology, microbiology and infectious disease, genetics, and prenatal/newborn screening. Commercially available tests for clinical applications are being developed for variants with diagnostic, prognostic, and therapeutic significance in specific disease types.
    Summary: The power of dPCR technology relies on the partitioning of the reactions and results in increased sensitivity and accuracy compared to qPCR. More recently, the sensitivity of dPCR has been applied to the detection of known variants in cell-free DNA and circulating tumor DNA. Future clinical applications of dPCR include liquid biopsy, treatment resistance detection, screening for minimal residual disease, and monitoring allograft engraftment in transplanted patients.
    MeSH term(s) Pregnancy ; Female ; Infant, Newborn ; Humans ; Polymerase Chain Reaction/methods ; Prenatal Diagnosis ; Nucleic Acids
    Chemical Substances Nucleic Acids
    Language English
    Publishing date 2024-01-02
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ISSN 2576-9456
    ISSN 2576-9456
    DOI 10.1093/jalm/jfad103
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Periductal Stromal Tumor of the Breast with a

    Anderson, Blaire / Marotti, Jonathan D / Lefferts, Joel A / Muller, Kristen E

    International journal of surgical pathology

    2023  Volume 31, Issue 8, Page(s) 1626–1631

    Abstract: The molecular pathogenesis of breast fibroepithelial tumors continues to be elucidated. Recently, highly ... ...

    Abstract The molecular pathogenesis of breast fibroepithelial tumors continues to be elucidated. Recently, highly recurrent
    MeSH term(s) Humans ; Female ; Phyllodes Tumor/diagnosis ; Phyllodes Tumor/genetics ; Phyllodes Tumor/pathology ; Mediator Complex/genetics ; Mediator Complex/metabolism ; Breast/pathology ; Breast Neoplasms/genetics ; Breast Neoplasms/pathology ; Mutation ; Fibroadenoma/pathology ; Telomerase/genetics
    Chemical Substances Mediator Complex ; TERT protein, human (EC 2.7.7.49) ; Telomerase (EC 2.7.7.49)
    Language English
    Publishing date 2023-02-23
    Publishing country United States
    Document type Case Reports ; Journal Article
    ZDB-ID 1336393-1
    ISSN 1940-2465 ; 1066-8969
    ISSN (online) 1940-2465
    ISSN 1066-8969
    DOI 10.1177/10668969231157306
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: Lessons Learned from Direct-to-Consumer Genetic Testing.

    Petersen, Lauren M / Lefferts, Joel A

    Clinics in laboratory medicine

    2020  Volume 40, Issue 1, Page(s) 83–92

    Abstract: The number of companies offering direct-to-consumer genetic tests is increasing. There is growing concern over whether direct-to-consumer genetic companies should be allowed to offer clinically relevant testing that has only been possible under medical ... ...

    Abstract The number of companies offering direct-to-consumer genetic tests is increasing. There is growing concern over whether direct-to-consumer genetic companies should be allowed to offer clinically relevant testing that has only been possible under medical care. Direct-to-consumer genetic testing can be incomplete, inaccurate, and inappropriate. The usefulness of such testing is extremely limited and it is unclear how well customers understand reported results. Research on the long-term impact of direct-to-consumer genetic testing is necessary to determine if stricter regulations regarding the performance of direct-to-consumer genetic testing are necessary.
    Language English
    Publishing date 2020-01-07
    Publishing country United States
    Document type Journal Article ; Review
    ZDB-ID 604580-7
    ISSN 1557-9832 ; 0272-2712
    ISSN (online) 1557-9832
    ISSN 0272-2712
    DOI 10.1016/j.cll.2019.11.005
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  4. Article ; Online: Development and validation of a molecular tool to predict pathologic complete response in esophageal adenocarcinoma.

    Qian, David C / Lefferts, Joel A / Zaki, Bassem I / Brickley, Elizabeth B / Jackson, Christopher R / Andrici, Juliana / Sriharan, Aravindhan / Lisovsky, Mikhail

    Diseases of the esophagus : official journal of the International Society for Diseases of the Esophagus

    2024  Volume 35, Issue 12

    Abstract: Pathologic complete response (pCR) to neoadjuvant chemoradiation for locally advanced esophageal adenocarcinoma (EAC) confers significantly improved survival. The ability to infer pCR may spare esophagectomy in some patients. Currently, there are no ... ...

    Abstract Pathologic complete response (pCR) to neoadjuvant chemoradiation for locally advanced esophageal adenocarcinoma (EAC) confers significantly improved survival. The ability to infer pCR may spare esophagectomy in some patients. Currently, there are no validated biomarkers of pCR. This study sought to evaluate whether a distinct signature of DNA copy number alterations (CNA) can be predictive of pCR in EAC. Pretreatment biopsies from 38 patients with locally advanced EAC (19 with pCR and 19 with pathologic partial/poor response) were assessed for CNA using OncoScan assay. A novel technique was employed where within every cytogenetic band, the quantity of bases gained by each sample was computed as the sum of gained genomic segment lengths weighted by the surplus copy number of each segment. A threefold cross-validation was used to assess association with pCR or pathologic partial/poor response. Forty patients with locally advanced EAC from The Cancer Genome Atlas (TCGA) constituted an independent validation cohort. Gains in the chromosomal loci 14q11 and 17p11 were preferentially associated with pCR. Average area under the receiver operating characteristic curve (AUC) for predicting pCR was 0.80 among the threefold cross-validation test sets. Using 0.3 megabases as the cutoff that optimizes trade-off between sensitivity (63%) and specificity (89%) in the discovery cohort, similar prediction performance for clinical and radiographic response was demonstrated in the validation cohort from TCGA (sensitivity 61%, specificity 82%). Copy number gains in the 14q11 and 17p11 loci may be useful for prediction of pCR, and, potentially, personalization of esophagectomy in EAC.
    MeSH term(s) Humans ; Treatment Outcome ; Esophageal Neoplasms/therapy ; Esophageal Neoplasms/drug therapy ; Adenocarcinoma/genetics ; Adenocarcinoma/therapy ; Esophagectomy ; Neoadjuvant Therapy/methods
    Language English
    Publishing date 2024-02-22
    Publishing country United States
    Document type Journal Article
    ZDB-ID 639470-x
    ISSN 1442-2050 ; 1120-8694
    ISSN (online) 1442-2050
    ISSN 1120-8694
    DOI 10.1093/dote/doac035
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article ; Online: Rare Activating BRAF Alteration Involving the β3-αC Kinase Domain in Ganglioglioma.

    Lin, Chun-Chieh / Lefferts, Joel A / Chan, Amy M / Zanazzi, George

    Journal of neuropathology and experimental neurology

    2021  Volume 80, Issue 9, Page(s) 887–889

    MeSH term(s) Adult ; Brain Neoplasms/diagnosis ; Brain Neoplasms/genetics ; Brain Neoplasms/metabolism ; Ganglioglioma/diagnosis ; Ganglioglioma/genetics ; Ganglioglioma/metabolism ; Humans ; Male ; Proto-Oncogene Proteins B-raf/genetics ; Proto-Oncogene Proteins B-raf/metabolism
    Chemical Substances BRAF protein, human (EC 2.7.11.1) ; Proto-Oncogene Proteins B-raf (EC 2.7.11.1)
    Language English
    Publishing date 2021-03-10
    Publishing country England
    Document type Case Reports ; Letter ; Research Support, N.I.H., Extramural
    ZDB-ID 3088-0
    ISSN 1554-6578 ; 0022-3069
    ISSN (online) 1554-6578
    ISSN 0022-3069
    DOI 10.1093/jnen/nlab013
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  6. Article ; Online: SNP-based chromosomal microarray characterization in a series of pure erythroid leukemia.

    Mindiola Romero, Andres E / Johnston, Michael A / Giffin, Justin T / Khan, Wahab A / Lefferts, Joel A / Loo, Eric Y

    Leukemia & lymphoma

    2022  Volume 63, Issue 8, Page(s) 2009–2012

    MeSH term(s) Humans ; Leukemia, Erythroblastic, Acute ; Myelodysplastic Syndromes
    Language English
    Publishing date 2022-04-03
    Publishing country United States
    Document type Letter ; Research Support, Non-U.S. Gov't
    ZDB-ID 1042374-6
    ISSN 1029-2403 ; 1042-8194
    ISSN (online) 1029-2403
    ISSN 1042-8194
    DOI 10.1080/10428194.2022.2057489
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  7. Article ; Online: Implementation of Reverse Transcriptase PCR Testing for Severe Acute Respiratory Syndrome Coronavirus 2 under the US Food and Drug Administration Emergency Use Authorization.

    Lefferts, Joel A / Allen, Samantha F / Steinmetz, Heather B / Tsongalis, Gregory J

    Clinical chemistry

    2020  Volume 67, Issue 2, Page(s) 434–435

    MeSH term(s) COVID-19/diagnosis ; COVID-19/virology ; COVID-19 Testing/methods ; Humans ; Reverse Transcriptase Polymerase Chain Reaction/methods ; SARS-CoV-2/isolation & purification ; United States ; United States Food and Drug Administration
    Language English
    Publishing date 2020-12-14
    Publishing country England
    Document type Letter
    ZDB-ID 80102-1
    ISSN 1530-8561 ; 0009-9147
    ISSN (online) 1530-8561
    ISSN 0009-9147
    DOI 10.1093/clinchem/hvaa278
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  8. Article ; Online: Squamous Cell Carcinoma in Hidradenitis Suppurativa Lesions Following Tumor Necrosis Factor α Inhibitors.

    Cooper, Shaun D / Cowdrey, Molly C E / Linos, Konstantinos D / Lefferts, Joel A / Basic, Katherina K

    Cutis

    2021  Volume 107, Issue 4, Page(s) E5–E7

    MeSH term(s) Carcinoma, Squamous Cell/chemically induced ; Hidradenitis Suppurativa/drug therapy ; Humans ; Male ; Middle Aged ; Tumor Necrosis Factor Inhibitors/adverse effects
    Chemical Substances Tumor Necrosis Factor Inhibitors
    Language English
    Publishing date 2021-06-07
    Publishing country United States
    Document type Case Reports ; Letter
    ZDB-ID 391840-3
    ISSN 2326-6929 ; 0011-4162 ; 0151-9522
    ISSN (online) 2326-6929
    ISSN 0011-4162 ; 0151-9522
    DOI 10.12788/cutis.0233
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  9. Article ; Online: Cross-reactivity of NRASQ61R antibody in a subset of Spitz nevi with 11p gain: a potential confounding factor in the era of pathway-based diagnostic approach.

    Parra, Ourania / Lefferts, Joel A / Tafe, Laura J / Gru, Alejandro A / Linos, Konstantinos

    Human pathology

    2021  Volume 112, Page(s) 35–47

    Abstract: The most recent World Health Organization classification for skin tumors (2018) categorizes melanomas and their precursor lesions, benign or intermediate, into nine pathways based not only on their clinical and histomorphologic characteristics but also ... ...

    Abstract The most recent World Health Organization classification for skin tumors (2018) categorizes melanomas and their precursor lesions, benign or intermediate, into nine pathways based not only on their clinical and histomorphologic characteristics but also on their molecular profile and genetic fingerprint. In an index case of a partially sampled atypical spitzoid lesion, which proved to be an 11p-amplified Spitz nevus with HRASQ61R mutation, we observed cross-reactivity with the NRASQ61R antibody (clone SP174). Overall, we assessed the status of HRAS and NRAS genes and their immunoreaction to NRASQ61R antibody in 16 cases of 11p-amplified Spitz nevi/atypical Spitz tumors. We also assessed the immunoexpression of NRASQ61R antibody in various malignancies with proven BRAFV600E, NRASQ61R, L or K, KRASQ61R and HRASQ61R, and HRASQ61R mutations and ALK+ Spitz lesions. Finally, we assessed the expression of PReferentially expressed Antigen in MElanoma (PRAME) immunohistochemistry in our 11p Spitz cohort. Three of 16 cases (3/16) harbored the HRASQ61R mutation and exhibited diffuse immunoreaction with the NRASQ61R antibody. All the cases in our cohort were negative for the NRASQ61R mutation. All NRASQ61R-, KRASQ61R-, and HRASQ61R-mutated neoplasms were positive for the antibody, further supporting the cross-reactivity between the RAS proteins. All the cases of our cohort were essentially negative for PRAME immunohistochemistry. In the era of pathway-based approach in the diagnosis of melanocytic neoplasms, the cross-reactivity between the NRASQ61R- and HRASQ61R-mutated proteins can lead to a diagnostic pitfall in the assessment of lesions with spitzoid characteristics.
    MeSH term(s) Adolescent ; Adult ; Aged ; Antibodies, Monoclonal/immunology ; Biomarkers, Tumor/analysis ; Biomarkers, Tumor/genetics ; Child ; Chromosomes, Human, Pair 11 ; Cross Reactions ; Diagnosis, Differential ; Female ; GTP Phosphohydrolases/genetics ; Humans ; Male ; Melanoma/diagnosis ; Melanoma/genetics ; Membrane Proteins/genetics ; Middle Aged ; Mutation ; Nevus, Epithelioid and Spindle Cell/diagnosis ; Nevus, Epithelioid and Spindle Cell/genetics ; Proto-Oncogene Proteins p21(ras)/genetics ; Skin Neoplasms/diagnosis ; Skin Neoplasms/genetics ; Young Adult
    Chemical Substances Antibodies, Monoclonal ; Biomarkers, Tumor ; Membrane Proteins ; GTP Phosphohydrolases (EC 3.6.1.-) ; NRAS protein, human (EC 3.6.1.-) ; HRAS protein, human (EC 3.6.5.2) ; Proto-Oncogene Proteins p21(ras) (EC 3.6.5.2)
    Language English
    Publishing date 2021-02-24
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural
    ZDB-ID 207657-3
    ISSN 1532-8392 ; 0046-8177
    ISSN (online) 1532-8392
    ISSN 0046-8177
    DOI 10.1016/j.humpath.2021.02.001
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  10. Article ; Online: Implementation of an Emergency Use Authorization Test During an Impending National Crisis.

    Lefferts, Joel A / Gutmann, Edward J / Martin, Isabella W / Wells, Wendy A / Tsongalis, Gregory J

    The Journal of molecular diagnostics : JMD

    2020  Volume 22, Issue 7, Page(s) 844–846

    Abstract: The laboratory response to the current severe acute respiratory syndrome coronavirus 2 pandemic may be termed heroic. From the identification of the novel coronavirus to implementation of routine laboratory testing around the world to the development of ... ...

    Abstract The laboratory response to the current severe acute respiratory syndrome coronavirus 2 pandemic may be termed heroic. From the identification of the novel coronavirus to implementation of routine laboratory testing around the world to the development of potential vaccines, laboratories have played a critical role in the efforts to curtail this pandemic. In this brief report, we review our own effort at a midsized, rural, academic medical center to implement a molecular test for the virus; and we share insights and lessons learned from that process, which might be helpful in similar situations in the future.
    MeSH term(s) Betacoronavirus/isolation & purification ; COVID-19 ; COVID-19 Testing ; Clinical Laboratory Techniques/standards ; Clinical Laboratory Techniques/statistics & numerical data ; Coronavirus Infections/diagnosis ; Coronavirus Infections/prevention & control ; Coronavirus Infections/virology ; Delivery of Health Care/organization & administration ; Emergencies ; Health Plan Implementation ; Humans ; Laboratories/legislation & jurisprudence ; Laboratories/standards ; Pandemics/prevention & control ; Pneumonia, Viral/diagnosis ; Pneumonia, Viral/prevention & control ; Pneumonia, Viral/virology ; SARS-CoV-2
    Keywords covid19
    Language English
    Publishing date 2020-05-14
    Publishing country United States
    Document type Journal Article ; Review
    ZDB-ID 2000060-1
    ISSN 1943-7811 ; 1525-1578
    ISSN (online) 1943-7811
    ISSN 1525-1578
    DOI 10.1016/j.jmoldx.2020.05.001
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