Heteroduplex PCR analysis of rearranged immunoglobulin genes for clonality assessment in multiple myeloma.

BACKGROUND AND OBJECTIVE: Molecular analysis by PCR of monoclonally rearranged immunoglobulin (Ig) genes can be used for diagnosis in B-cell lymphoproliferative disorders (LPD), as well as for monitoring minimal residual disease (MRD) after treatment. This technique has the risk of false-positive results due to the "background" amplification of similar rearrangements derived from polyclonal B-cells. This problem can be resolved in advance by additional analyses that discern between polyclonal and monoclonal PCR products, such as the heteroduplex analysis. A second problem is that PCR frequently fails to amplify the junction regions, mainly due to somatic mutations frequently present in mature (post-follicular) B-cell lymphoproliferations. The use of additional targets (e.g. Ig light chain genes) can avoid this problem. DESIGN AND METHODS: We studied the specificity of heteroduplex PCR analysis of several Ig junction regions to detect monoclonal products in samples from 84 MM patients and 24 patients with B cell polyclonal disorders. RESULTS: Using two distinct VH consensus primers (FR3 and FR2) in combination with one JH primer, 79% of the MM displayed monoclonal products. The percentage of positive cases was increased by amplification of the Vlamda-Jlamda junction regions or kappa(de) rearrangements, using two or five pairs of consensus primers, respectively. After including these targets in the heteroduplex PCR analysis, 93% of MM cases displayed monoclonal products. None of the polyclonal samples analyzed resulted in monoclonal products. Dilution experiments showed that monoclonal rearrangements could be detected with a sensitivity of at least 10(-2) in a background with >30% polyclonal B-cells, the sensitivity increasing up to 10(-3) when the polyclonal background was

Evaluating the objective structured long examination record for nurse education

An objective structured long examination record (OSLER) is a modification of the long-case clinical examination and is mainly used in medical education. This study aims to obtain nursing students' views of the OSLER compared with the objective structured clinical examination (OSCE), which is used to assess discrete clinical skills. A sample of third-year undergraduate nursing students (n=21) volunteered to participate from a cohort of 230 students. Participants undertook the OSLER under examination conditions. Pre-and post-test questionnaires gathered the students' views on the assessments and these were analysed from a mainly qualitative perspective. Teachers' and simulated patient views were also used for data triangulation. The findings indicate that the OSLER ensures more holistic assessment of a student's clinical skills and particularly essential skills such as communication, and that the OSLER, together with the OSCE, should be used to supplement the assessment of clinical competence in nursing education.