79 research outputs found
A comparison of the distribution of pneumococcal types in systemic disease and the upper respiratory tract in adults and children
The serotype distribution of 874 strains of Streptococcus pneumoniae was determined in relation to patients' age and to frequency of isolation from systemic disease. Types 14 and 18, in pre-school children, and types 1, 4, 7, 8 and 12 in patients over 5 years of age were significantly associated with systemic disease whereas type 23 in pre-school children, and type 6 in older patients was associated with upper respiratory tract carriage. No significant difference was found in the incidence of other types in systemic disease compared to upper respiratory tract carriage. Fifteen diagnostic pneumococcal antisera (to types 1, 3, 4, 6, 7, 8, 9, 11, 12, 14, 17, 18, 19, 22 and 23) sufficed for typing 87% of strains
Measurement of Epstein-Barr virus DNA load using a novel quantification standard containing two EBV DNA targets and SYBR Green I dye
<p>Abstract</p> <p>Background</p> <p>Reactivation of Epstein-Barr virus (EBV) infection may cause serious, life-threatening complications in immunocompromised individuals. EBV DNA is often detected in EBV-associated disease states, with viral load believed to be a reflection of virus activity. Two separate real-time quantitative polymerase chain reaction (QPCR) assays using SYBR Green I dye and a single quantification standard containing two EBV genes, Epstein-Barr nuclear antigen-1 (EBNA-1) and BamHI fragment H rightward open reading frame-1 (BHRF-1), were developed to detect and measure absolute EBV DNA load in patients with various EBV-associated diseases. EBV DNA loads and viral capsid antigen (VCA) IgG antibody titres were also quantified on a population sample.</p> <p>Results</p> <p>EBV DNA was measurable in ethylenediaminetetraacetic acid (EDTA) whole blood, peripheral blood mononuclear cells (PBMCs), plasma and cerebrospinal fluid (CSF) samples. EBV DNA loads were detectable from 8.0 × 10<sup>2 </sup>to 1.3 × 10<sup>8 </sup>copies/ml in post-transplant lymphoproliferative disease (n = 5), 1.5 × 10<sup>3 </sup>to 2.0 × 10<sup>5 </sup>copies/ml in infectious mononucleosis (n = 7), 7.5 × 10<sup>4 </sup>to 1.1 × 10<sup>5 </sup>copies/ml in EBV-associated haemophagocytic syndrome (n = 1), 2.0 × 10<sup>2 </sup>to 5.6 × 10<sup>3 </sup>copies/ml in HIV-infected patients (n = 12), and 2.0 × 10<sup>2 </sup>to 9.1 × 10<sup>4 </sup>copies/ml in the population sample (n = 218). EBNA-1 and BHRF-1 DNA were detected in 11.0% and 21.6% of the population sample respectively. There was a modest correlation between VCA IgG antibody titre and BHRF-1 DNA load (rho = 0.13, p = 0.05) but not EBNA-1 DNA load (rho = 0.11, p = 0.11).</p> <p>Conclusion</p> <p>Two sensitive and specific real-time PCR assays using SYBR Green I dye and a single quantification standard containing two EBV DNA targets, were developed for the detection and measurement of EBV DNA load in a variety of clinical samples. These assays have application in the investigation of EBV-related illnesses in immunocompromised individuals.</p
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