Human rhinovirus : development of an experimental disease program and detection in tissue employing in situ hybridisation and a polymerase chain reaction

Bardin, Philip Greyling (1994) Human rhinovirus : development of an experimental disease program and detection in tissue employing in situ hybridisation and a polymerase chain reaction. University of Southampton, Doctoral Thesis.

Abstract

Human Rhinoviruses cause the majority of common colds and are etiologically linked with changes in lower airway physiology and asthma exacerbations. To investigate this association in more depth, we have developed a program of experimental volunteer inoculation and obtained detailed measurements of symptoms, viral shedding and nasal albumin levels as well as biopsy material from both the nose and lower airways.

Atopic individuals had amplified cold symptoms accompanied by increased nasal albumin levels. Significant increases in airway responsiveness were noted in atopic but not normal volunteers; decreases in peak expiratory flow occurred equally in both groups in 30% of cases. Bronchoscopy and nasal biopsy procedures were well tolerated.

In situ hybridisation was developed to detect intracellular Rhinovirus employing oligonucleotide biotinylated probes. Detection of hybrids could be enhanced by labeling on both ends of the probe and the virus was observed in the nasal epithelial cells of 3/6 subjects with colds. A polymerase chain reaction assay developed to detect Rhinovirus in tissues was used to evaluate deaths caused by asthma; no evidence was found to implicate the virus.

The validated experimental cold program and molecular techniques that have been developed will expedite definition of the link between the common cold, atopy and asthma.

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