Journal of Clinical Laboratory Analysis最新文献

Background: The longitudinal study was conducted over the initial 2 years of the COVID-19 pandemic, spanning from June 2020 to December 2022, in healthcare workers (HCWs) of the Thomayer University Hospital. A total of 3892 blood samples were collected and analyzed for total nucleocapsid (N) antibodies. The aim of the study was to evaluate the dynamics of N antibodies, their relationship to the PCR test, spike (S) antibodies, interferon-gamma, and prediction of reinfection with SARS-CoV-2.

Methods: Blood collections were performed in three rounds, along with questionnaires addressing clinical symptoms of past infection, PCR testing, and vaccination. Antibody measurements included total N antibodies (Roche Diagnostics) and postvaccination S antibodies (Euroimmun). Cellular immunity was tested by interferon-gamma release assay (Euroimmun).

Results: At the end of the study, 35.9% of HCWs were positive for N antibodies, and 39.5% of HCWs had either known PCR positivity or N antibodies or both. Ten percent of participants had no knowledge of a COVID-19 infection and 35% of positive individuals exhibited no symptoms. The values of positive antibodies decrease over a period of 6 months to 1 year, depending on the initial value, and their dynamics are highly variable. The study also demonstrated that the highest levels of spike antibodies and interferon-gamma occur during so-called hybrid immunity.

Conclusion: Nucleocapsid antibodies proved valuable in monitoring SARS-CoV-2 infection dynamics, and they may detect cases of SARS-CoV-2 infection missed by PCR tests. The study identified distinct patterns in antibody dynamics and protection of hybrid immunity during reinfection.

Objectives: To determine the association between IGF2BP2 polymorphisms and type 2 diabetes mellitus.

Methods: This study involved 422 individuals, 214 diabetes mellitus cases, and 208 healthy controls. The PCR-RFLP technique was used to determine the genotype of the IGF2BP2 gene for the SNPs rs4402960 (G>T) and rs1470579 (A>C).

Results: The results showed that the C allele of the rs1470579 variant and the T allele of the rs440960 variant were significantly associated with type 2 diabetes mellitus group (p = 0.0029 and 0.0001) as reported for both alleles, respectively. High frequencies of haplotypes TA and TC were observed in type 2 diabetes patients.

Conclusion: The present study showed that IGF2BP2 rs1470579 and rs4402960 polymorphism were significantly associated with the increased risk of T2DM in the Saudi Arabian population and presented a genetic model to screen the high-risk individuals with further validations.

Background: Low-temperature cryopreservation is a common method for scientific research and clinical sample preservation when utilizing flow cytometry. In flow cytometry data analysis, traditional manual "gating" is susceptible to past experience and faces the challenge of manual subjective bias, time-consuming, and multidimensional data analysis. With the development of algorithms, the advantages of dimensionality reduction and clustering in result analysis are gradually becoming more prominent.

Methods: Flow cytometry was used to detect the effects of cryopreservation and freeze-thaw cycle on T-cell subsets, and to analyze the data using automated algorithmic analysis and conventional manual "gating" methods.

Results: The results showed that the number and viability of cells decreased slightly after one freeze-thaw within 2 weeks of cryopreservation, and there was no significant change in the subpopulation proportions and spatial locations by both analysis methods. The changes were significant with the increase of cryopreservation time and freeze-thaw cycle, which may be due to changes in the molecular conformation of the maker as a result of cryopreservation.

Conclusion: The results indicate that both analysis methods have reached similar conclusions, but the repeatability and objectivity of automated algorithmic analysis have compensated for the uncertainty brought about by the subjective discretization of traditional manual "gating." In addition, the automated algorithmic analysis more intuitively highlights the spatial positional variations in the relationships between cell populations.

Background: Inductively coupled plasma mass spectrometry (ICP-MS) is widely used for the accurate measurement of minerals. However, its application to serum essential mineral measurement has not been fully evaluated. The present study aimed to assess the performance of ICP-MS for serum minerals by comparing its measurements to those obtained using standard quantification methods.

Methods: Cross-sectional data were collected from 282 participants from a single facility in Japan. Serum concentrations of eight key minerals, namely sodium, potassium, calcium, phosphorus, magnesium, iron, zinc, and copper, measured via ICP-MS and standard methods were compared using Passing-Bablok regression and Bland-Altman plots.

Results: All minerals, except phosphorus, exhibited good agreement with standard methods, with more stable regression coefficients observed for minerals with greater interindividual variability. After systematically filtering outliers, the mean relative errors were approximately -3% for sodium, potassium, calcium, and magnesium; +5% for iron; 0% for zinc; and -19% for copper. The outliers for iron were primarily due to mild hemolysis, whereas those for zinc were largely attributed to nonhemolysis factors. For phosphorus, the serum total phosphorus concentration measured using ICP-MS was approximately 3.5 times higher than the serum inorganic phosphorus concentration measured using standard methods, with a weak correlation observed between the two methods.

Conclusion: This study provides a practical foundation for future research. Understanding ICP-MS characteristics will facilitate the development of new approaches in clinical diagnostics.

Background: Breast cancer (BC) is an aggressive tumor. Circulating tumor cells (CTCs) are a potential biomarker for the prognosis of cancer patients. This study aimed to explore the prognostic significance of CTCs in patients with BC.

Methods: Retrospectively, 108 BC patients were collected between January 2011 and December 2019, while 10 patients with benign nodules were included as controls. CTCs with different phenotypes of patients were isolated using CanPatrol and tricolor RNA in situ hybridization (RNA-ISH) methods. Estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor-2 (HER-2) levels were measured by immunohistochemistry (IHC). The progression-free survival (PFS) was calculated using the Kaplan-Meier method. Independent risk factors for BC recurrence were determined by Cox proportional risk regression analysis.

Results: The higher the cancer stage (p = 0.00), the higher the ki-67 expression level (p < 0.01), and the lower the histologic grade (p < 0.01), the higher the number of CTCs. The PFS of patients with high CTCs was shorter than that of patients with low CTCs (p < 0.05). Total CTCs (≥ 6) and positive mesenchymal CTCs (MCTCs) were also associated with recurrence and metastasis.

Conclusions: Total CTCs in BC patients have an independent influence on PFS reduction. Higher total CTCs and MCTCs in peripheral blood are biomarkers for predicting the prognosis of BC patients. HER-2 high expression is also associated with the prognosis of the disease.

Background: Research on heart injury caused by COVID-19 is limited to large observational and retrospective cohort studies using imaging or pathological data. Reported changes in the levels of myocardial markers in severe diseases have been limited, with few studies on mild infections. The effects of COVID-19 on cardiac function and changes in myocardial marker levels have not yet been reported.

Methods: We analyzed data from outpatient blood samples collected at Beijing Anzhen Hospital during the 2022 COVID-19 outbreak and used the same periods in 2020 and 2021 as controls, focusing on changes in routine blood tests, coagulation, myocardial markers, and other blood indices in patients with palpitations.

Results: The number of patients with palpitations increased by 4.87-fold during the COVID-19 mass outbreak in 2022. The indices of myocardial damage did not show any symptom-related increases but decreased within the normal range. The proportion of patients with palpitations whose D-dimer and fibrinogen/fibrin degradation product (FDP) values exceeded the reference ranges increased, as did the numbers of neutrophils, monocytes, and platelets. In this retrospective analysis, we found little change in the myocardial markers in patients with mild COVID-19.

Conclusions: In patients with mild COVID-19, attention should be diverted from detecting myocardial markers to changes in coagulation test results, focusing on the levels of coagulation indices to improve circulation and prevent thrombosis.

Background: HPV plays a key role in the development of cervical cancer. This study aims to investigate the prevalence of HPV genotypes in patients with Squamous cell carcinoma (SSC) and Adenocarcinoma (ADC) at NORI cancer Hospital Pakistan, with the aim of improving screening and prevention strategies.

Method: Cervical scrapings were collected from 129 diagnosed cervical cancer patients. HPV typing was performed using a real-time PCR assay and sequencing.

Result: Among the patients, 73.6% (90/129) were HPV positive. Proportion of HPV positivity was observed within each group. The highest incidence of HPV was observed in the 50-60 years age group (80.9%), followed by the 40-to-50-year group (75.8%). The positivity rate declined in the 60-to-70-year-old (63.6%) and further in the 70-80 years (62.5%). Eight different HPV subtypes were identified, with HPV 16 being the most prevalent (80.0%), followed by HPV 18 (9.5%), HPV 45 in 2 (2.1%), HPV 31 in 1 (1.1%), HPV 35 in 1 (1.1%), HPV 59 in 1 (1.1%), HPV 66 in 1 (1.1%), and HPV 89 in 1 (1.15). Histologically, 89.2% of the cases were Squamous cell carcinoma (SCC) and 10.8% were Adenocarcinomas. In SCC patients, HPV 16 was found in 80.9% of cases, while in Adenocarcinoma patients, HPV 16 was detected in 66.7% of cases.

Conclusion: The prevalence of high-risk HPV types, both vaccine and non-vaccine, targeted in Pakistan highlights the urgent need for widespread screening and vaccination programs. Tailored public health strategies are essential to effectively reduce cervical cancer rates and mortality.

Background: To establish a dual immunoassay based on inductively coupled plasma mass spectrometry (ICP-MS) with stable element labeling antibodies for the simultaneous detection of alpha-fetoprotein (AFP) and prostate-specific antigen (PSA) in serum and evaluate its performance and clinical sample validation.

Methods: The immunoassay system based on the double antibody sandwich method was established using magnetic beads as solid-phase carriers and rare earth elements europium (Eu) and samarium (Sm) as element tags. The test conditions were optimized. According to the Clinical Laboratory Standards Institute (CLSI) guidelines and the "Guidelines for Performance Validation of Clinical Chemistry Quantitative Testing Procedures," performance validation was conducted using the established method, followed by clinical sample validation.

Results: The LOD and AMR for AFP were 0.76 ng/mL and 1.5-1200 ng/mL, respectively; the LOD and AMR for PSA were 0.41 ng/ mL and 0.5-250 ng/ mL, respectively. The intra-batch precision for AFP and PSA was all less than 3.33%, and the inter-batch precision for AFP and PSA was all less than 4.97%. The recoveries of AFP and PSA were 95.08%-104.18% and 97.65%-100.72%, respectively. The correlation coefficients between the established and the CLIA methods were r = 0.9930 for AFP and r = 0.9852 for PSA.

Conclusions: We have developed a dual immunoassay method based on ICP-MS, which can quantitatively detect AFP and PSA in serum within a single analysis. This method exhibits good precision, a wide linear range, and high specificity, meeting the requirements for clinical sample detection and holding potential for clinical application.

Background: Thromboelastography (TEG) and coagulation tests can be used to detect hypercoagulability to assess thrombus formation. This study explored the association between TEG and coagulation in evaluating disease severity in elderly patients with lower extremity arteriosclerotic occlusive disease (LEASO), aimed to provide surgical treatment guideline.

Methods: We retrospectively analyzed the clinical characteristics, laboratory biomarkers, TEG and coagulation parameters of 233 elderly LEASO patients treated between 2020 and 2023. Among them, 86 underwent surgical amputation, 51 received vascular intervention, and the remaining were treated conservatively. Differences in TEG and coagulation among the three groups were assessed using Spearman's correlation. Multivariate logistic regression and receiver operating characteristic curves analyzed the relationships among TEG, fibrinogen (FIB), and D-dimer (D-D) levels for surgical evaluation.

Results: Inflammatory factors, platelet counts, and Fontaine stages III-IV differed significantly between the surgery and conservative groups (p < 0.05). The surgery group had higher α-angle, maximum amplitude (MA), coagulation comprehensive index (CI), FIB, and D-D, whereas lower clotting time (K) compared to the conservative group (p < 0.05), correlated with a lower ankle brachial index (ABI), indicating more severe clinical presentation. Spearman's analysis identified positive associations between α-angle, MA with FIB and D-D levels in surgical patients. Area under curve analysis indicated that combining MA, α-angle, FIB, and D-D could enhance accuracy in evaluating surgical necessity in LEASO.

Conclusion: In elderly LEASO patients, TEG and coagulation analysis revealed a positive association between thrombus intensity and disease severity. Increased MA, α-angle, FIB, and D-D levels serve as predictors for surgical treatment necessity in LEASO.

Background: SAP2 is closely associated with the pathogenicity and drug resistance of Candida albicans (C. albicans). Our study aimed to construct C. albicans with SAP2 overexpression (pRB895-SAP2-SC5314) to explore the influence of SAP2 on the adhesion of C. albicans and predict the interaction between magnolol and Sap2 by molecular docking.

Methods: The recombinant plasmid pRB895-SAP2 with high SAP2 expression was prepared using a plasmid extraction kit and transformed into C. albicans strain SC5314 using an improved lithium acetate conversion method to construct PRB895-SAP2-SC5314. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was performed to detect the expression of adhesion-related genes in the different strains. Molecular docking and visual analysis of magnolol and Sap2 were performed using the CB-DOCK2 platform.

Results: Compared with the control SC5341 and SC5341 transfected with pRB895, SAP2 expression was significantly higher in the pRB895-SAP2-SC5314 strain (p < 0.05). Based on the sequencing and mapping results, the pRB895-SAP2-SC5314 strain was successfully prepared. SAP2 overexpression significantly downregulated ALS1 expression (p < 0.05), whereas ALS3, TEC1, HOG1, PHR1, and TUP1 expression was downregulated in C. albicans (p < 0.05). The optimal docking result for magnolol and Sap2 protein was -8.1 kcal/mol of vina score, which was considered good docking.

Conclusions: SAP2 overexpression may strengthen the adhesion and pathogenicity of C. albicans, and magnolol may act as an Sap2 inhibitor that affects the adhesion of C. albicans.