Laboratory medicine最新文献

Objective: The diagnosis and prognosis of hepatocellular carcinoma (HCC) present significant challenges in clinical practice. This study aimed to evaluate the clinical utility of tumor abnormal protein (TAP), Prothrombin induced by vitamin K absence-II (PIVKA-II), and alpha-fetoprotein (AFP) in diagnosing HCC as well as to investigate their prognostic significance in patients with HCC undergoing transarterial chemoembolization.

Methods: A total of 93 HCC patients were enrolled and 101 healthy individuals served as controls. Fresh venous blood samples were collected, and TAP, PIVKA-II, and AFP levels were measured by chemiluminescence immunoassay.

Results: Significant differences in TAP, PIVKA-II, and AFP levels were found between HCC patients and healthy individuals. The combined assay of TAP, AFP, and PIVKA-II showed better diagnostic performance for HCC. Patients who underwent transarterial chemoembolization and achieved complete response (CR) had lower levels of prechemotherapy serum TAP, AFP, and PIVKA-II. There are significant differences in levels of TAP, AFP, and PIVKA-II between CR and partial response (PR), CR and stable disease (SD), and CR and progressive disease (PD).

Conclusion: Combined detection of TAP, PIVKA-II, and AFP has better diagnostic performance for HCC. Higher levels of prechemotherapy serum TAP, AFP, and PIVKA-II are significantly associated with poor clinical chemoresponse.

Background: CALR mutation analysis is routinely used to diagnose BCR/ABL1-negative myeloproliferative neoplasms. The 2 most common CALR mutations are a 52-base pair (bp) deletion and a 5-bp insertion, which account for approximately 85% of cases.

Methods: To evaluate our new microfluidic chip assay, we tested CALR mutant and wild-type specimens that were previously analyzed using conventional methods at a reference laboratory. Samples included EDTA-anticoagulated peripheral blood and bone marrow specimens, air dried bone marrow aspirate smears, and formalin-fixed, paraffin-embedded bone marrow sections. CALR exon 9 was PCR amplified using 2 previously published primer pairs and a third unique primer pair designed for our new assay. Amplicons were sized using microfluidic chip analysis.

Results: Concordance with the reference method was 100% (42/42). Intra-run and inter-run reproducibility were also 100% (3/3 and 3/3, respectively). The limit of detection was confirmed to be 6% mutant alleles.

Conclusion: We determined that the microfluidic chip assay to detect CALR exon 9 mutations was acceptable for clinical use. Compared with the conventional method, the microfluidic analysis assay benefits from a streamlined workflow, faster turnaround, and a smaller instrument footprint.

Progesterone is a steroid hormone primarily associated with pregnancy. A simple, rapid, and reliable high-performance liquid chromatography (HPLC) method has been developed and validated for the quantification of progesterone in human plasma. The method consists of a simple liquid-liquid extraction of progesterone and internal standard (estriol) from human plasma using a mixture of hexane and diethyl ether. The chromatographic determination of progesterone was performed using an acetonitrile-water (70:30, v/v) mobile phase with a C18 reversed-phase column. The method achieved an extraction recovery of greater than 96.4% from spiked plasma samples. Intra- and inter-day precision were generally acceptable, with relative SD% less than ≤6.60% and accuracy (relative error %) better than 3.64%. The developed and validated method was used to successfully quantify progesterone levels in plasma samples collected from women during the third trimester of pregnancy. Furthermore, a statistical comparison was conducted between progesterone concentrations in plasma samples obtained from 2 groups of pregnant women: group 1 (n = 9) at 30-35 weeks and group 2 (n = 9) at 36-41 weeks. The developed and validated HPLC method described in this study enables the successful determination of progesterone in human plasma, offering advantages such as shorter analysis time, simplicity, cost-effectiveness, and potential routine use during pregnancy.

Objective: Aldosterone synthase deficiency (ASD) is a rare autosomal recessive inherited disease with an overall clinical phenotype of failure to thrive, vomiting, severe dehydration, hyperkalemia, and hyponatremia. Mutations in the CYP11B2 gene encoding AS are responsible for the occurrence of ASD. Defects in CYP11B2 gene have only been reported in a limited number of cases worldwide. Due to this potential life-threatening risk, a comprehensive hormonal investigation followed by genetic confirmation is essential for the clinical management of offspring.

Methods: We report a case of a newborn who was found to have persistent hyponatremia, hyperkalemia, low aldosterone level, raised renin levels, normal cortisol, and normal 17 hydroxyprogesterone level, suggesting the diagnosis of isolated ASD.

Results: Genetic report was suggestive of isolated ASD caused by a novel base pair deletion in exon 3, homozygous CYP11B2 variant (chr8:g.142915123_142915125del; depth: 124x d) (p.Lys175del; ENST00000323110.2). After initial steps of rehydration and salt restoration, the child was started on oral tablet fludrocortisone. The child responded well and showed a good gain in growth and development.

Discussion: We elaborate on the biochemical and genetic work-up performed and describe potential pitfalls in CYP11B2 sequencing due to its homology to CYP11B1.

Background: The kappa-free light chain (κFLC) index has shown its value in detecting the intrathecal synthesis of immunoglobulins. We aimed to evaluate the diagnostic performance of the κFLC index for multiple sclerosis (MS) and compare different algorithms proposed in the literature to optimize its use for our population.

Methods: Based on the results of the oligoclonal bands (OCBs) and κFLC index of 255 patients with suspected MS different optimization strategies were evaluated, for which the optimal κFLC index cut-off thresholds were calculated.

Results: The best diagnostic performance was achieved by using a reflexive algorithm, in which OCBs are only performed according to the κFLC index result. With a single cut-off (κFLC index = 7.9), an accuracy of 92.2% was obtained (sensitivity = 92.4%, specificity = 92%) with an OCB performance rate of 58.1%. When applying 2 cut-offs (κFLC index = 4.2 and 13), the accuracy was the same (92.2%, sensitivity = 89.6%, specificity = 94%), but the OCB performance rate dropped to 29.4%.

Conclusion: The 2-step strategy proposed with κFLC determination followed by OCB analysis in the borderline cases appears to be the most suitable solution, further optimized by adjusting the decision thresholds to 4.2 < κFLC index < 13, resulting in high accuracy and the most saving of OCBs.

Objective: The aim of the study was to compare the cost and clinical impact of repeating BioFire FilmArray gastrointestinal (GI) and respiratory (RP) panel assays with 3 vs 4 pathogen targets positive.

Method: We analyzed 12,027 GI and RP panels to evaluate our retesting policy, which retested panels with 3 or more detected pathogens (3-pathogen protocol) compared with the manufacturer's 4-pathogen (4-pathogen protocol) recommendation. We compared the retesting results, calculated the cost implications, and reviewed the clinical impact on antibiotic prescriptions and patient outcomes.

Results: Retesting with our 3-pathogen protocol revealed that 81% (39/48) of GI and 76% (26/34) of RP panels had identical results, whereas 19% (9/48) of GI and 24% (8/34) of RP panels showed discrepancies on retesting. The additional cost incurred by our protocol compared with the manufacturer's protocol was $9820.32. There was no evidence that our more stringent policy affected antibiotic prescription or clinical outcomes.

Conclusion: Our more stringent 3-pathogen protocol for retesting panels did not improve patient management compared with the manufacturer's 4-pathogen protocol but resulted in unnecessary costs and increased the risk of depleting testing kits during supply shortages. Consequently, we adopted the manufacturer's suggestions, highlighting the need to balance clinical rigor with cost-effectiveness in laboratory testing protocols.

Background: Gene polymorphisms of rearranged during transfection (RET) and its ligand neurturin (NRTN) are one of the focus of studies in the investigation of cancer pathogenesis, invasion, and metastasis. In this study, we aimed to examine the possible risk of breast cancer between RET G691S, L769L, S904S, and NRTN IVSI-663 polymorphisms and to evaluate serum NRTN, brain-derived neurotrophic factor (BDNF), matrix metalloproteinase (MMP)-2, MMP-9, and focal adhesion kinase (FAK) levels.

Methods: The study consists of 110 breast cancer patients and 110 controls. Polymorphisms were detected by the polymerase chain reaction method from study groups whole blood.

Results: The NRTN IVSI-663 polymorphism in G allele has been found to be 1.54 fold increased the risk of breast cancer, however AA genotype has been found 0.43 fold decreased the risk of breast cancer (P < .05, P < .05, respectively). Study groups showed a similar profile for RET G691S, L769L, S904S allele frequencies and genotype distributions (P > .05). In the patient group, significant increase in serum NRTN and FAK levels and decrease in MMP-2 and MMP-9 levels were found (P < .05, P < .05, P < .05, P < .05, respectively).

Discussion: In summary that increased breast cancer risk with the G allele in NRTN gene IVSI-663 polymorphism, as well as the increased serum NRTN and FAK levels, will contribute to the diagnosis, prognosis and determination of new treatment strategies.

Background: One possible way to improve accuracy of blood culture Gram stain analyses is increasing the concentration of organisms on the slide prepared from the blood culture broth.

Methods: From each positive blood culture bottle, 1 direct smear, a 1-drop concentrated preparation, and 1 cytospin/cytofuge preparation were Gram stained and evaluated. There were 2 evaluators who ranked the 3 preparations from most to fewest organisms seen. Each preparation was also scored as acceptable or unacceptable for both organism and background stain quality.

Results: The 1-drop slide exhibited the highest concentration of organisms compared with both the cytospin and direct smear but presented frequent difficulties with interpretation of Gram staining acceptability with both the background and the organisms. Although cytospin preparations are known to concentrate liquid specimens, the current study found no enrichment of microorganisms over the direct smear preparation. The cytospin was, however, advantageous for enhancing organism morphology by creating a monolayer of elements and reducing background artifacts.

Conclusion: The 1-drop preparation is simple, inexpensive, and effective at increasing organism concentration for analysis, making it a good option when processing culture bottles with low organism loads, but more investigation into methods of improving stain quality is necessary.

Specimen hemolysis is a frequent finding when blood is collected from neonates. This produces artificially high results for some analytes, such as potassium. Testing samples for electrolytes using point-of-care (POC) blood gas analyzers is convenient and facile. However, unlike testing that is conducted on serum or plasma from a central laboratory, detection of hemolysis using POC analyzers cannot currently be achieved. As described in these cases, the presence of hemolysis can produce ambiguities and delays in the diagnosis and management of neonates.

Background: The use of xylene substitutes is becoming more common in the setting of micrographic surgery frozen tissue section staining, and dermatologic surgeons need to be aware of possible undesirable delayed effects of using these agents and the possibility of modifying H&E staining protocols to prevent delayed fading. This report demonstrates an undesirable outcome of using an isoparaffinic aliphatic hydrocarbon as a xylene substitute, implementation of a quality improvement intervention to eliminate frozen section slide fading in the setting of micrographic surgery tissue processing, and recommendations for the modification of protocol when using a xylene substitute.

Clinical and laboratory information: Frozen section slides processed with xylene and xylene substitute were analyzed by histotechnicians, a dermatopathologist, and a micrographic surgery surgeon at 1-week and 1-month intervals. The use of a standard H&E protocol resulted in zero stains fading when using xylene as a clearing agent, but delayed fading when using a xylene substitute.

Discussion: Using an isoparaffinic aliphatic hydrocarbon as a xylene substitute can lead to excess water carryover, which may result in delayed hematoxylin fading in micrographic surgery tissue staining, so using this xylene substitute likely requires modification to the dehydration phase and tap water immersion phase to prevent fading.