Laboratory medicine最新文献

Introduction: The prognosis of acute myeloid leukemia is stratified by genetic abnormalities; however, the detection sensitivity varies by method. The KIT D816 mutation is frequently found in core binding factor leukemia and is associated with a poor prognosis. In this study, we aimed to investigate the performance of multiplex droplet digital polymerase chain reaction (ddPCR) for detecting KIT D816 mutations and propose the practical mutation analysis method for clinical laboratory testing.

Methods: We evaluated the detection limit of ddPCR using mixed probes for HEX-labeled wild-type and FAM-labeled mutations (D816V, D816Y, and D816H) by analyzing plasmid mixtures containing these sequences. We compared the frequency of KIT mutations detected by direct sequencing and ddPCR in 20 patients with core binding factor leukemia.

Results: Multiplex ddPCR successfully discriminated between mutation types based on plot positions on a 2-dimensional map, with a detection limit of 0.1%. The frequency of D816 mutations was 42.5% using ddPCR and 20% using direct sequencing. Most patients with KIT D816 mutation require hematopoietic stem cell transplantation for chimeric gene clearance.

Discussion: Amplitude-based multiplex ddPCR efficiently provides sensitive and accurate genotyping of KIT D816 and has potential applications for other genetic abnormalities.

Introduction: Patients with hematologic malignancies who undergo immunosuppressive therapies such as chimeric antigen receptor T-cell (CAR-T) therapy are at high risk of prolonged SARS-CoV-2 infection due to impaired humoral immunity. Treatment options remain limited, with variable efficacy, in such settings.

Methods: We describe a 21-year-old man with Down syndrome and B-cell acute lymphoblastic leukemia complicated by B-cell aplasia following CD19-directed CAR-T therapy. The patient developed COVID-19 and experienced persistent symptomatic infection, with high viral load and prolonged reverse transcriptase-polymerase chain reaction (RT-PCR) positivity for more than 7 months.

Results: Despite multiple courses of remdesivir and extended weekly infusions of COVID-19 convalescent plasma (CCP), the patient remained viremic and intermittently symptomatic. Anti-SARS-CoV-2 immunoglobulin G titers were detectable only toward the latter time frame of treatment, and passive antibody therapy with CCP was insufficient for viral clearance. Ultimately, compassionate use of monoclonal antibody (mAb) therapy (casirivimab and imdevimab) was granted. Following administration, the patient achieved viral clearance for the first time, with resolution of symptoms and persistently negative RT-PCR findings for 8 months of available follow-up thereafter.

Discussion: This case illustrates the limitations of CCP in patients with prolonged SARS-CoV-2 infection and highlights the effectiveness of mAbs in achieving viral clearance in severely immunocompromised hosts. It supports targeted use of mAb therapy in select high-risk populations and reinforces the importance of specific passive immunotherapy strategies (when available) for the management of viremia in immunodeficient patients.

Introduction: For several years, concerns about perfluoroalkyl and polyfluoroalkyl substances (PFAS) have been growing, and clinical laboratories may have to perform a growing number of PFAS analyses. The use of serum tubes without a separator gel is currently recommended for the quantification of PFAS due to the concern that the compounds may adsorb to the gel. The impact of gel adsorption on the accuracy of the results, however, has not been evaluated.

Methods: Aliquots from a pool of blood spiked with PFAS were stored in gel-free clot activator tubes (CATs) and gel-containing BD SST II Advance tubes (serum separator tubes) for 2, 8, and 24 hours. The CATs and serum separator tubes were collected from 15 volunteers under typical sampling conditions. Concentrations of 16 PFAS were analyzed using liquid chromatography-tandem mass spectrometry, and the percentage of change in PFAS levels between both tube types was computed.

Results: Results showed minimal changes (<5%) for most PFAS within 24 hours, except for long-chain perfluorosulfonates (including perfluorooctane sulfonate), which seemed to exhibit adsorption of the gel. In samples from volunteers, the observed changes were statistically significant for perfluorooctane sulfonate (P <.001).

Discussion: Based on our analysis, we recommend using CATs to avoid PFAS underestimation.

Introduction: Hepatocellular carcinoma (HCC) diagnosis requires a combination of elevated ɑ-fetoprotein (AFP) levels and liver imaging. We evaluated the role of prothrombin induced by vitamin K antagonist-II (PIVKA-II) in patients with suspected HCC.

Methods: A retrospective analysis was conducted on adult Indian patients with suspected HCC who had elevated AFP and PIVKA-II levels and had undergone radiologic investigation.

Results: Among 178 suspected cases, 144 (80.99%) were diagnosed with HCC; the median age of these 144 patients was 62 years (IQR, 54-68 years). The median AFP and PIVKA-II levels were 14.68 ng/mL (IQR, 3.83-146.50 ng/mL) and 354.50 ng/mL (IQR, 49.41-4293.40 ng/mL), respectively. The sensitivity and specificity were 52.08% and 88.24% for AFP (≥10 ng/mL) and 80.56% and 32.35% for PIVKA-II (≥40 ng/mL). The false positivity rate of PIVKA-II was 67.60%. PIVKA-II showed moderate correlation (r = 0.413) with HCC size and was more reliable than AFP for HCC lesions larger than 5 cm (area under the curve = 0.731 vs 0.627). PIVKA-II levels were lower following therapy for HCC (682.98-122.50 ng/mL; P < .001) than AFP levels (15.05-6.50 ng/mL; P = .30). Among AFP-negative patients with HCC, 68.10% had elevated PIVKA-II levels.

Discussion: With its higher false positivity rate, PIVKA-II may not be a reliable marker for HCC, although it may be useful in larger HCC lesions and in AFP-negative patients. Establishing a newer cutoff value for PIVKA-II among the Indian population may improve its specificity in the diagnosis and management of HCC.

The American Society for Clinical Pathology Board of Certification Research and Development Committee has undertaken regular surveys of Medical Laboratory Science (MLS) education programs. Results of previous surveys were reported in 2019 and 2022. The purpose of these recurring surveys is to support MLS programs in their educational mission by providing information about current issues and trends that may affect program resources and MLS education, including data for grant applications, funding requests, and strategic planning. Program-related topics covered in the surveys included program director and faculty education and certification requirements, program structure and duration, student grade point averages and acceptance data, clinical site numbers and status, and faculty numbers and vacancies.

Introduction: Colorectal cancer (CRC) is known to have an association with circular RNAs (circRNAs) and immune checkpoint factors. This study sought to examine the expression levels of hsa_circ_0000284 and hsa_circ_0004771 molecules and the programmed cell death 1 protein (PD-1) and ecto-5'-nucleotidase (CD73) immune checkpoints in the peripheral blood of patients with CRC as well as the ratios of circular to linear forms of homeodomain-interacting protein kinase 3 (HIPK3) and nuclear receptor interacting protein 1 (NRIP1).

Methods: Real-time polymerase chain reaction (PCR) and flow cytometry were used to assess quantitatively the expression level of circRNAs, CD73, and PD-1 in blood samples from patients with CRC and healthy control individuals. The expression of CD73 and PD-1 molecules was analyzed using FlowJo software, and the expression levels of circRNAs, CD73, and PD-1 were calculated with real-time PCR analysis.

Results: Real-time PCR analysis revealed a statistically significant increase in the linear form of hsa_circ_0004771 (linNRIP1) and PD-1 gene expression in patients' blood compared with control individuals. In addition, the circHIPK3:linHIPK3 and circNRIP1:linNRIP1 ratios are statistically significantly higher in patients than in healthy control individuals. The flow cytometry assessment indicated a statistically significant increase in PD-1 on the surface of lymphocytes and monocytes and an increase in CD73 on the granulocytes of patients compared with the healthy control individual.

Discussion: Based on these findings, hsa_circ_0000284, hsa_circ_0004771, PD-1, and CD73 were statistically significantly increased in our cancer group. If further research were done, these blood markers could potential be biomarkers for CRC progression.

Introduction: This study investigated the association between thyroid disorders and breast cancer.

Methods: A retrospective case-control study was conducted using electronic health record data from an academic health care system. This study included 300 female patients 18 years of age and older with a diagnosis of breast cancer and 300 control individuals with no cancer history. Statistical analysis included calculation of prevalence and odds ratios for association, ꭓ2 for categorical variables, and mean (SD) and median for age.

Results: Patients with breast cancer were, on average, 58 years old, statistically significantly older than patients without cancer (P < .001). Patients were 6 times more likely to have thyroid disorders (P < .001), including hypothyroidism (P < .001), than women who did not have cancer. Patients with triple-negative breast cancer had 4 times higher odds of being diagnosed with thyroid disorders overall (P < .001) and hypothyroidism (P < .001) compared with control individuals. Moreover, patients with estrogen receptor-positive/progesterone receptor-positive breast cancer were 15 times more likely to develop the autoimmune disorder Hashimoto thyroiditis compared with controls, which was statistically significant (P = .026).

Discussion: This study highlighted a strong association between breast cancer and thyroid disorders, particularly indicating a higher prevalence of hypothyroidism among individuals with breast cancer.

Introduction: The coexistence of primary thyroid lymphoma (PTL) and papillary thyroid carcinoma (PTC) is extremely rare, especially in young populations. Therefore, it is extremely important to understand and master the clinical manifestations, diagnosis, treatment, and prognosis of these diseases.

Methods: This article reports a case of a 29-year-old female patient with recurrent right neck swelling and pain with fever and progressive enlargement of the thyroid gland.

Results: The imaging examination results showed goiteromegaly and a mass. Subsequent surgical resection was performed, and postoperative pathology confirmed the coexistence of PTC and PTL. The patient then received 8 cycles of chemotherapy plus targeted therapy. During follow-up, the tumor disappeared, and the disease was in complete remission.

Discussion: It is extremely rare for a young patient with recurrent neck pain and fever to have PTC and PTL simultaneously. We report this case to provide a basis for the diagnosis and treatment of the coexistence of PTC and PTL to improve clinicians' understanding of these coexisting diseases in young people to avoid misdiagnosis and missed diagnosis.

Introduction: Prostate cancer is one of the most commonly diagnosed cancers in men worldwide, and early detection is essential for improving survival. Current diagnostic methods, such as prostate-specific antigen tests and biopsies, have limitations, emphasizing the need for noninvasive biomarkers.

Methods: Proteomics analysis was performed on urine samples from patients with prostate cancer and healthy individuals to identify differentially expressed proteins. Publicly available datasets, including the National Center for Biotechnology Information Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA), were analyzed to validate gene expression patterns and their association with survival outcomes.

Results: Proteomics analysis identified 18 statistically significantly altered proteins in patients with prostate cancer, including reduced expression of ciliary neurotrophic factor receptor (CNTFR). Validation with GEO and TCGA datasets confirmed lower CNTFR expression in prostate cancer tissues than in normal tissues. Reduced CNTFR expression was associated with poorer overall survival and disease-free survival.

Discussion: CNTFR is a promising noninvasive diagnostic biomarker for prostate cancer. Its reduced expression in urine and tissues, along with its association with poor prognosis, highlights its potential for improving prostate cancer diagnosis and outcomes through noninvasive methods.

Introduction: Early-stage diagnosis is crucial for the best prognosis in breast cancer. Here, our objective was to compare serum tumor protein p53 (TP53) levels in an Egyptian population with benign breast disease vs breast cancer to investigate the protein's utility in early detection and intervention, alongside existing tumor-related markers, including carcinoembryonic antigen (CEA), cancer antigen 125 (CA125), and CA15.3.

Methods: We enrolled 231 patients in this study: 146 with breast cancer, 50 with benign conditions, and 35 without breast pathology (healthy control individuals). The serum level of TP53, CEA, CA125, and CA15.3 was quantitated using an enzyme-linked immunosorbent assay. Data were examined using the t test, Spearman correlation, and receiver operating characteristic curve analysis.

Results: Patients with breast cancer showed clinically significant higher serum levels of TP53 than did individuals with benign conditions and healthy control individuals. Elevated TP53 levels were correlated with advanced tumor stage (P <.001) and high-tumor grade (P <.001). TP53 effectively distinguished between benign disease and early-stage breast cancer, with an area under the curve of 0.88, 73.6% sensitivity, and 78.0% specificity. Multinomial logistic regression analysis showed that combining TP53 with other tumor-related markers improved early breast cancer detection. Both TP53 and CA125 remained significant predictors in this analysis. A new function, the PCA-Index, which combines concentrations of TP53 and CA125, was developed to enhance early detection in early stages (T1-T2) and low grades (1-2) with notable results: areas under the curve of 0.96 and 0.98, a specificity of 90.0%, and sensitivities of 85.7% and 91.7%, respectively.

Discussion: The combined detection of TP53 and CA125, the PCA-Index, has important clinical value for early detection of breast cancer with a high degree of sensitivity and specificity.