Biomedical subjects

A M Roque Afonso

Publications and source records attributed to A M Roque Afonso.

3 recordsLinked to original sources

PubMed2000

Performance of an automated system for quantification of hepatitis C virus RNA.

The Amplicor HCV Monitor test for quantitative determination of serum or plasma hepatitis C virus (HCV) RNA was modified recently and introduced onto the Cobas Amplicor instrument to automate fully amplification, detection and calculation of results. This new version (v2.0) was evaluated in a routine diagnostic laboratory. The sensitivity and reproducibility were assessed on well-characterized panels (Eurohep) and clinical samples. HCV RNA levels measured by the v2.0 Monitor test were about 1log(10) higher than those detected by the previous version test, and genotypes 1 and 3 were quantified with equal sensitivity. Within the linear dynamic range of 10(3) to 10(6) copies/ml, the coefficients of variation for both intra- and inter-assay reproducibility ranged from 1.9 to 2.95%. This test system was found to be a reliable and labor saving assay for the quantification of HCV RNA.

Automation↗

PubMed1999

Nonrandom distribution of hepatitis C virus quasispecies in plasma and peripheral blood mononuclear cell subsets.

The existence of an extrahepatic reservoir of hepatitis C virus (HCV) is suggested by differences in quasispecies composition between the liver, peripheral blood mononuclear cells, and serum. We studied HCV RNA compartmentalization in the plasma of nine patients, in CD19(+), CD8(+), and CD4(+) positively selected cells, and also in the negatively selected cell fraction (NF). HCV RNA was detected in all plasma samples, in seven of nine CD19(+), three of eight CD8(+), and one of nine CD4(+) cell samples, and in seven of eight NF cells. Cloning and sequencing of HVR1 in two patients showed a sequence grouping: quasispecies from a given compartment (all studied compartments for one patient and CD8(+) and NF for the other) were statistically more genetically like each other than like quasispecies from any other compartment. The characteristics of amino acid and nucleotide substitutions suggested the same structural constraints on HVR1, even in very divergent strains from the cellular compartments, and homogeneous selection pressure on the different compartments. These findings demonstrate the compartmental distribution of HCV quasispecies within peripheral blood cell subsets and have important implications for the study of extrahepatic HCV replication and interaction with the immune system.

Amino Acid Sequence↗

PubMed1998

A control programme for MRSA (methicillin-resistant Staphylococcus aureus) containment in a paediatric intensive care unit: evaluation and impact on infections caused by other micro-organisms.

Methicillin-resistant Staphylococcus aureus (MRSA) is increasingly reported as a hospital-acquired pathogen in intensive care units (ICUs). The inconsistent application of hygiene measures by healthcare workers accounts largely for the epidemic dissemination of such resistant strains. The efficacy of a control programme to prevent spread of MRSA was assessed in our paediatric ICU (PICU) from April 1992 to December 1995. Patients initially had weekly MRSA cultures taken from samples of anterior nares and perineum, but from January 1994, cultures were also obtained upon admission. Immediately after notification, all MRSA carriers were isolated. Education of hospital staff was an essential component of our programme. Nosocomial infection rates were recorded retrospectively in 1992 and 1993, and prospectively in 1994 and 1995. Incidence rates between 'pre-programme' and 'programme' periods were compared. The rate of MRSA infection decreased from 5.9-0.8/1000 Patient-Days (PD), (P < 10(-7). MRSA carriage also decreased from 34-2% (P < 10(-9) and the ratio of MRSA to all S. aureus fell from 71-11% (P < 10(-4). The decrease in the global incidence of infection from 20.1-13.9/1000 PD (P = 0.002) was due only to the decrease in MRSA infection. However, between 1994 and 1995, there was a significant increase in the number of transplant patients despite a constant patient/nurse ratio. The nosocomial infection rates caused by other micro-organisms decreased among the transplant patients from 64.8-33.2/1000 transplanted PD (P = 0.009) between 1994 and 1995. At the same time, we observed a slight increase of infections in non-transplanted patients, which may have been due to the effect of increased overall workload on those patients who were supposed to have fewer nosocomial risk factors. We conclude that implementation of infection control measures directed towards limiting person-to-person spread was effective in controlling high MRSA infection rates in a PICU, but it is important to allow enough time for staff to carry out hygiene practices thoroughly.

Child↗