In March 2003, a novel coronavirus (SARS-CoV) was discovered in association with cases of severe acute respiratorysyndrome (SARS). The sequence of the complete genome of SARS-CoV was determined, and the initial characterization of the viral genome is presented in this report. The genome of SARS-CoV is 29,727 nucleotides in length and has 11 open reading frames, and its genome organization is similar to that of other coronaviruses. Phylogenetic analyses and sequence comparisons showed that SARS-CoV is not closelyrelated to anyof the previouslycharacterized coronaviruses.

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The polymerase chain reaction (PCR) assay is a powerful tool for diagnosis of infectious, genetic, and neoplastic diseases (1). However, standard methods of PCR product analysis-gel electrophoresis followed by ethidium bromide staining, and Southern blot hybridization to confirm product identity, remain tedious and time-consuming, preventing applied transfer of these procedures to clinical laboratories. Several solid-phase colorimetric assays that capture denatured PCR products on probes bound to nylon membranes (2, 3) or to microtiter plates (4, 5) offer improved convenience, but may lack optimal sensitivity due to the tendency of the denatured PCR product strands to reassociate and exclude oligonucleotide probes, and steric interference from the solid support that impedes hybridization. In some cases, colorimetric detection is improved by creating single-strand PCR products through asymmetric PCR that can associate with bound probes without interference (6). Unfortunately, asymmetric PCR is notoriously difficult to reproduce, and does not lend itself to automation. To circumvent these problems, we describe a simple, highly specific, nonradioactive microtiter plate format for detection of

This is a case of a 36-year-old gentleman with haemophilia A who was presented with an acute atraumatic soft tissue swelling in the right thigh. Open biopsy was performed with the resultant diagnosis of a synovial cell sarcoma. Although the clinical findings were nonspecific they could easily have been found in a bleeding haemophilic pseudotumour. The findings reported on MRI scan initially were highly consistent with those present in patients with mild haemophilia. An important part of orthopaedic management in haemophilia is concerned with intraarticular and intramuscular bleeding. Haematomas are common and sarcomas are rare. However the absence of trauma should alert the clinician to the possibility that the abnormality may represent haemorrhage into a tumour and not just haematoma, even in a haemophilic patient.

K-ras point mutations are often detected in part of the lung carcinomas. For the validation of a highly sensitive and rapid assay for known point mutations, Point-EXACCT (Biochim Biophys Acta 1998; 1379:42–52), we analyzed 89 non-small cell lung carcinomas and compared the results with two sequencing methods. No point mutations were found with double-stranded sequencing. Single-stranded sequencing detected six patients positive for K-ras codon 12. When Point-EXACCT was used, K-ras codon 12 mutations were detected in 8 of 52 patients with squamous cell carcinomas, 10 of 29 patients with adenocarcinomas, and 3 of 8 patients with large cell carcinomas. The finding of K-ras mutations in squamous cell carcinomas is explained by the high sensitivity of the method. Therefore, Point-EXACCT may be applicable to detection of those alterations occurring at a low frequency among an excess of cells with wild-type DNA.

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