High-resolution melting of PCR amplicons with the DNA dye LCGreen I was recently introduced as a homogeneous, closed-tube method genotyping that does not require probes or real-time PCR. We adapted this system to genotype single-nucleotide polymorphisms (SNPs) after rapid-cycle (12 min) small (</=50 bp).Engineered plasmids were used study all possible SNP base changes. In addition, clinical protocols for factor V (Leiden) 1691G>A, prothrombin 20210G>A, methylenetetrahydrofolate reductase...

High-resolution melting was recently introduced as a technique to genotype single-nucleotide polymorphisms (SNPs) within small amplicons (1). This closed-tube method (including rapid-cycle PCR) can be completed in <15 min and does not require real-time PCR instruments (2), allele-specific (3), or fluorescently labeled oligonucleotides (4)(5)(6). The process is made possible by heteroduplex-detecting DNA dyes that used at saturating concentrations without inhibiting (7). Wild-type homozygous...

This study investigates differences in expression of the cell cycle/growth activation markers p53, p16, and p27, their relationship with nerve sheath proliferation among plexiform neurofibromas (PNF), NF1-related non-NF1 MPNSTs different histologic grades between benign-appearing malignant areas associated PNFs. Formalin-fixed, paraffin-embedded archival tissue from PNFs were immunostained using avidin-biotin-complex method antibodies to S-100 protein (S-100), Leu7 (CD57), CD34, Mib-1,...

IAN is an R package that addresses the challenge of integrating, analyzing and interpreting high-throughput "omics" data, using a multi-agent artificial intelligence (AI) system. leverages popular pathway regulatory datasets (KEGG, WikiPathways, Reactome, GO, ChEA) STRING database for protein-protein interactions to perform standard enrichment analysis. The individual results are then used generate insightful summaries, each datasets, large language model (LLM) through architecture. These...

The measurement of multiple antigens in a single sample poses clinical and methodological challenges. Here we describe the validation multiplexed sandwich enzyme-linked immunosorbent assay (ELISA) array (microELISA) nine antigens. tested simultaneously were: α-fetoprotein (AFP), prostate specific antigen (PSA), carcinoembryonic (CEA), cancer 125 (CA 125), CA 15-3, 19-9, β-human chorionic gonadotropin (β-hCG), luteinizing hormone (LH), follicle stimulating (FSH). At least 44 samples were for...

Two methods for closed-tube single nucleotide polymorphism (SNP) genotyping without labeled probes have become available: unlabeled probe and amplicon melting. Unlabeled melting assays were compared using 5 SNPs: human platelet antigens 1, 2, 5, 15 a C>T variant located 13,910 base pairs (bp) upstream of the lactase gene. LCGreen Plus (Idaho Technology, Salt Lake City, UT) was used as saturating DNA dye. data readily interpretable accurate all lengths tested. Five targets that ranged in size...

Abstract Large B‐cell lymphoma with IRF4 rearrangement is a provisional entity in the 2017 World Health Organization classification. In order to characterize these lymphomas children from United States, FISH and immunohistochemical stains were performed on 32 follicular diffuse large (DLBCL) Children's Oncology Group studies. Two DLBCLs (6%) had rearrangements, one involving ileocecal valve another tonsil cerebrospinal fluid. Both cases strong, IRF4/MUM1 staining, which may be pathologic...

ABSTRACT Determination of the hepatitis C virus (HCV) genotype for infected patients increasingly has become accepted as standard care. Genotype assignment helps in assessing disease prognosis and assists establishing appropriate duration treatment. The great genetic diversity HCV, with 11 major genotypes &gt;70 subtypes, contributes to technical difficulty testing. While “gold standard” testing is nucleic acid sequencing, a variety hybridization assays, including line probe assay, have been...

Introduction: Detection of MYC translocations using fluorescence in situ hybridization (FISH) is important the evaluation lymphomas, particular, Burkitt lymphoma and diffuse large B-cell lymphoma. Our aim was to validate a digital FISH capture imaging system for detection 8q24 LSI-MYC (a break-apart probe) 8;14 translocation IGH-MYC fusion probe). Materials Methods: probe evaluated tissue sections from 35 patients. forty Sections were processed analyzed traditional methods. slides then...

ABSTRACT The genotype of the infecting hepatitis C virus (HCV) helps determine patient's prognosis and duration treatment. Heteroduplex mobility analysis (HMA) is a rapid, inexpensive method for genotyping HCV that does not require sequencing. We developed an HMA uses temperature gradient capillary electrophoresis (TGCE) to differentiate genotypes. A 56-bp region 5′ untranslated (UTR) was conserved within yet whose sequence differed between genotypes amplified HMA-TGCE analysis. amplicons...

The eosin-5'maleimide (EMA) binding test has been studied extensively for the detection of hereditary spherocytosis (HS). Its performance characteristics have compared to NaCl-based or glycerol lysis-based red cell osmotic fragility tests and cryohemolysis. HS samples are also better identified when both mean channel fluorescence (MCF) EMA relative controls coefficient variation (CV) analyzed.We looked at 65 normal including 30 adults 25-65 years old 35 newborns 12 cases. In addition MCF CV,...

Paroxysmal nocturnal hemoglobinuria (PNH) is a rare acquired disorder characterized by increased complement-mediated lysis of erythrocytes (RBCs) because low/absent glycophosphatidylinositol (GPI) anchors numerous cell surface proteins.Rare event analysis was applied to 120 million RBCs (12 normal individuals) and 102 (102 establish reference range verify methodology for analysis. Patient PNH testing (n = 10,984) performed over 47 months using the 2010 consensus guidelines with CD59-PE...

Neurofibromatosis type 1 (NF1) is a common autosomal dominant genetic disorder that caused by mutation in the NF1 gene. Hallmark characteristics include dermal neurofibromas, café-au-lait spots, and learning disabilities. In approximately 25% of cases, plexiform or peripheral nerve sheath tumors (PNSTs) involve large segments root, can form, which small percentage become malignant (MPNST). Most MPNSTs are composed spindled neoplastic cells, they resemble other spindle-cell sarcomas,...

Summary Introduction Detection of chromosomal translocations in formalin‐fixed paraffin‐embedded ( FFPE ) leukemic samples is important for confirmation histopathological findings, classification, prognostication, and therapeutic decisions. Herein, we aim to determine whether digital expression profiling could detect identified by RT ‐ PCR , FISH and/or karyotyping. Methods RNA was extracted from 28 bone marrow specimens 19 patients diagnosed with leukemia. Eight were translocation t(9;22)...

Context.— B-cell lymphomas exhibit balanced translocations that involve immunoglobulin loci and result from aberrant V(D)J recombination, class switch or somatic hypermutation. Although most of the breakpoints in occur defined regions, those partner genes vary; therefore, it is unlikely 2 independent clones would share identical both partners. Establishing whether a new lesion patient with history lymphoma represents recurrence process can be relevant. Polymerase chain reaction (PCR)–based...

Plasminogen activator inhibitor type 2 (PAI-2) is a member of the serine protease (SERPIN) superfamily and forms stable complexes with urokinase plasminogen (uPA). uPA can be found on cell surface attached to its specific receptor (uPAR), allowing for controlled degradation extracellular matrix by activation into plasmin. The aim this study was evaluate if PAI-2 could also detected surface, providing means regulating activity uPA.Intact or permeabilized lines human peripheral blood...