Prevalence of prothrombin G20210A, factor V G1691A (Leiden), and methylenetetrahydrofolate reductase (MTHFR) C677T in seven different populations determined by multiplex allele-specific PCR.

Individuals belonging to six racial groups (African American, Asian Indian, Caucasian, Hispanic, Korean, Native American), and a seventh group comprised of referred patients with thrombosis were genotyped for the prothrombin G20210A mutation, the factor V G1691A (Leiden) mutation, and the methylenetetrahydrofolate reductase (MTHFR) C677T mutation by multiplexed allele-specific PCR. The prothrombin 20210A and factor V 1691A allele frequencies in the thrombosis patients, 3.2% and 9.

Detection of BCR/ABL RNA transcripts using the polymerase chain reaction is highly predictive for relapse in patients transplanted with unrelated marrow grafts for chronic myelogenous leukaemia.

Relapse after allogeneic bone marrow transplantation (BMT) for chronic myelogenous leukaemia (CML) is thought to result from residual leukaemia cells which survive the intensive conditioning regimen and are not eradicated by donor-derived immune effector cells capable of mediating a graft-versus-leukaemia (GVL) effect. Early relapse can be detected using highly sensitive assays such as the polymerase chain reaction (PCR) which have been shown to have predictive value for subsequent relapse in selected patient populations. The validity of PCR for predicting CML relapse in unrelated marrow transplant recipients where the GVL effect appears to be augmented due to increased HLA disparity between donor and recipient, however, has not been well defined.

The sensitivity of allele-specific polymerase chain reaction can obviate concern of maternal contamination when fetal samples are genotyped for immune cytopenic disorders.

Objective: Fetuses at risk for immune cytopenic disorders can be identified by molecular genotyping assays. To better understand the impact of maternal contamination on genotyping results, the levels of contamination that are routinely encountered during prenatal testing of fetal samples and the sensitivity of allele-specific polymerase chain reaction in detecting paternal alloalleles were examined.

Study Design: Reconstitution experiments were performed to define the sensitivity of allele-specific polymerase chain reaction assays.

Determination of neutrophil antigen gene frequencies in five ethnic groups by polymerase chain reaction with sequence-specific primers.

Background: The granulocyte antigens NA1 and NA2 are the two recognized allelic forms of Fc gamma receptor IIIB. These antigens are clinically relevant, because they are the most frequent targets of neutrophil antibodies in alloimmune neonatal neutropenia, transfusion-related acute lung injury, and chronic benign autoimmune neutropenia of infancy.

Study Design And Methods: A genotyping assay for NA1 and NA2 using polymerase chain reaction with sequence-specific forward and reverse oligonucleotide primers has been developed and validated.

Validation of multiplex polymorphic STR amplification sets developed for personal identification applications.

Polymorphic short tandem repeat (STR) loci, which typically consist of variations in the number of 3-7 base pair repeats present at a site, provide an effective means of personal identification. Typing can be accomplished by amplification of genomic DNA using the polymerase chain reaction (PCR) and locus-specific primers, separation of amplified alleles using gel electrophoresis and their display using silver staining or fluorescent detection. Primers for several STR loci can be combined in a single multiplex reaction so typing of multiple loci can be accomplished rapidly and with less DNA than required if each locus were analyzed separately.

Genotyping of KEL1 and KEL2 of the human Kell blood group system by the polymerase chain reaction with sequence-specific primers.

Background: Kell is a major antigenic system in human red cells, with more than 20 identified antigens. KEL1 and KEL2 are two opposing low- and high-frequency alleles. Immunization to KEL1 is clinically significant, because anti-KEL1 can cause severe reactions to transfusion of incompatible blood, as well as hemolytic disease of the newborn.

Use of unrelated marrow grafts compensates for reduced graft-versus-leukemia reactivity after T-cell-depleted allogeneic marrow transplantation for chronic myelogenous leukemia.

The effect of donor/recipient histocompatibility on relapse in patients receiving T-cell-depleted (TCD) grafts for chronic myelogenous leukemia (CML) was evaluated. Specifically, we sought to determine whether TCD results in an attenuation of the graft-versus-leukemia (GVL) effect on recipients of unrelated marrow grafts similar to that observed in HLA-identical sibling marrow transplantations. This question was addressed by comparative analysis of the relapse rates in marrow grafts who otherwise received identical preparative regimens and graft-versus-host disease (GVHD) prophylaxis schedules (T-cell depletion with T10B9 monoclonal antibody and complement plus posttransplant cyclosporine) and by serial molecular analyses using the polymerase chain reaction (PCR) to detect the bcr/abl RNA transcript in patients transplanted with unrelated donor grafts.

Replication of plasmid DNA in fertilized Xenopus eggs is sensitive to both the topology and size of the injected template.

The behavior of various plasmid templates was examined following their microinjection into fertilized eggs of the frog Xenopus laevis using an assay that permits the examination of both replicated and unreplicated plasmids in single eggs. Our results show that both the size and the topology of the template drastically affect the fate of the injected plasmid. Only a small proportion of injected monomeric supercoiled plasmids underwent replication during 6 h of incubation, although not all injected cells supported replication.