Multiplexed, ligation-dependent probe amplification for rapid and inexpensive HLA-DQB1 allelotyping.

Many effective options exist to accurately type DNA for human leukocyte antigen (HLA) alleles. However, most of the existing methods are excessively costly in terms of overall monetary costs, DNA requirements, and proprietary software. We present a novel assay capable of resolving heterozygous HLA-DQB1 allelotypes at two digits, with even greater specificity for the HLA-DQB1*06 allele family, by using the multiplexed ligation-dependent probe amplification technology. This assay provides more specific allele data than genome-wide analysis and is more affordable than sequencing, making it a useful intermediate for researchers seeking to accurately allelotype human DNA samples.

Computed tomographic scan can be used for surgical decision making in zone II penetrating neck injuries.

BACKGROUND: Controversy surrounds the evaluation of zone II penetrating neck injuries. Current literature supports mandatory exploration or selective management. Computed tomographic (CT) scanning provides high-resolution images that are used for trauma in other body regions. The purpose of this study is to prospectively evaluate the utility of CT scanning in the evaluation of zone II penetrating neck injuries. METHODS: From July 1998 to November 1999, 14 stable patients were studied who sustained zone II penetrating neck injuries. All patients had a physical examination, infusion CT scan of the neck, and an operative exploration. Before surgery, the trauma surgeon evaluated the CT scan and interpreted it as demonstrating either "high" or "low" probability for significant injury. Surgical findings were compared with the surgeon's preoperative interpretation of the CT scan. RESULTS: Three of 14 patients had five significant injuries. All these patients had high probability of injury CT scans, with four of the five injuries specifically diagnosed by CT scan. Sensitivity, specificity, positive predictive value, and negative predictive value were 100%, 91%, 75%, and 100% (p < 0.02), respectively. CONCLUSION: This small prospective study demonstrates that zone II penetrating neck injuries can be accurately evaluated by CT scan. In addition, the CT scan can be used for surgical decision making. This will eliminate the need for mandatory exploration and limit the role of angiography, esophagography, and endoscopy in zone II penetrating neck injuries.

Real-time reverse transcription polymerase chain reaction detection and quantification of t(1;19) (E2A-PBX1) fusion genes associated with leukaemia.

A real-time reverse transcription polymerase chain reaction (RT-PCR) method is described that enabled the detection and quantification of E2A-PBX1 fusion gene transcripts associated with t(1;19). The method was highly reproducible and offered exceptional sensitivity at 5 fg of fusion transcript per reaction, without the need for a nested PCR primer design. To illustrate the usefulness of this new technology the E2A-PBX1 fusion gene transcript expression level for several human leukaemia cell lines that are positive and negative for cytogenetically detectable t(1;19) was determined. The RCH-ACV had a threefold higher expression of E2A-PBX1 transcripts (600 transcripts per cell) than the other t(1;19) positive 697 (150 transcripts per cell). The only other cell line with detectable E2A-PBX1 was CEM, but the level of expression was < 1 transcript per cell.

A fast and simple method to determine the clonal relationship among human T-cell lymphocytes.

For the analysis of mutation in human T-cell lymphocytes, it is crucial to determine the clonal relationship between isolated mutants, particularly if they harbour identical mutations. Here we report an efficient method to determine the clonal relationships between these cells. The method is based on the analysis of restriction fragment length polymorphisms of a polymerase chain reaction amplified, rearranged T-cell receptor gamma-gene. As few as 600 cells are sufficient, regular agarose gels can be used for the separation of the restriction fragments, no radioactive label is required, and results can be obtained in 2 days.