Failure to diagnose recent hepatitis C virus infections in London injecting drug users.

Hepatitis C virus (HCV) infection results in chronic liver disease in a substantial proportion of those infected. Most new cases of HCV infection in the UK are associated with intravenous drug use. It is important to identify these infections because of the implications for the future health of the individuals concerned and for the control of further spread of infection.

The mcrA gene as an alternative to 16S rRNA in the phylogenetic analysis of methanogen populations in landfill.

Inferred amino acid sequences of the methyl coenzyme-M reductase (mcrA) gene from five different methanogen species were aligned and two regions with a high degree of homology flanking a more variable region were identified. Analysis of the DNA sequences from the conserved regions yielded two degenerate sequences from which a forward primer, a 32-mer, and a reverse primer, a 23-mer, could be derived for use in the specific PCR-based detection of methanogens. The primers were successfully evaluated against 23 species of methanogen representing all five recognized orders of this group of Archaea, generating a PCR product between 464 and 491 bp.

Multi-center European evaluation of HIV testing on serum and saliva samples.

Objective: to evaluate the reliability of HIV antibody testing on saliva.

Design: matched serum and saliva samples were collected from both seronegative (n = 344) and seropositive (n = 125) individuals in five European countries. Duplicate saliva samples collected with Omni-Sal devices provided by Saliva Diagnostic System (SDS) were pooled before analysis.

Differential diagnosis of HTLV-I and HTLV-II infections by restriction enzyme analysis of 'nested' PCR products.

A 'nested' polymerase chain reaction (PCR) assay is described which is capable of detecting single copies of human T-cell lymphotropic virus (HTLV) in genomic DNA extracted from peripheral blood mononuclear cells (PBMCs). A single set of 'nested' oligonucleotide primers, based on the highly conserved tax/rex region of the viral genome, was able to detect both HTLV-I and HTLV-II proviral sequences in clinical samples of diverse geographical origins, from the United States, Great Britain, Japan, the Caribbean, Italy, Greece, Iraq and West Africa. Rapid discrimination between HTLV-I and HTLV-II infections was achieved by restriction enzyme analysis of unpurified second-round PCR products, even in those cases in which serological assays had failed to provide a definitive result.

Comparison of ELISA, SPACE, and electron microscopy for the routine diagnosis of rotavirus infection.

Previous studies on the serological diagnosis of rotavirus infection have utilised locally produced antibodies. In this study we have compared two commercially produced assays, an ELISA (Rotazyme, Abbott) and a newly developed assay--solid phase aggregation of coupled erythrocytes (SPACE) (Wellcome Research Laboratories), with electron microscopy (EM). The SPACE test appeared less sensitive than EM.

Virus diarrhoea associated with pale fatty faeces.

Steatorrhoea was a significant feature in an outbreak of rotavirus gastroenteritis which affected adults and infants in hospital. Fat globules or fatty acid crystals were obvious by light microscopy (LM) in faeces from 14 of 25 patients examined. Ten of the fatty stools and two of the remainder were very pale.

Use of the ultracentrifuge vertical rotor in the detection of rubella-specific IgM on a sucrose density gradient.

A comparison was made of the performance of swing-out and vertical ultracentrifuge rotors in the detection of rubella-specific IgM on a sucrose density gradient. Tests were performed on 30 sera, of which 11 were found to contain rubella-specific IgM by both methods. The centrifugation time for the swing-out rotor was 16 hours at 35,000 rpm.

Rapid adenovirus typing by immunoelectron microscopy.

A rapid method of typing adenoviruses by immunoelectron microscopy is discribed. This emphasizes the value of an electron microscope in diagnostic virology, especially when a rapid result is required in epidemiology.