Further Observations on the Use of Organ Cultures in the Study of Acute Respiratorytract Infections

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Abstract
ALTHOUGH many viruses are known to cause acute respiratory-tract infections, the proportion of illnesses that can be diagnosed in the laboratory remains disappointingly small-about 30 per cent. The introduction of organ cultures of human embryonic trachea and nose (Tyrrell and Bynoe, 1965; Hoorn, 1966) and the discovery of the human respiratory coronaviruses (Tyrrell and Bynoe ; Hamre and Procknow, 1966) renewed hope that the virological investigation of such infections would be more profitable and the conclusions drawn from epidemiological studies less speculative. A comparison of the standard tissue-culture methods with organ cultures for the isolation of viruses from patients with acute respiratory infections (Higgins, Ellis and Woolley, 1969) showed that, although some viruses undetected by the standard methods could be isolated in organ cultures, both systems were necessary for maximum efficiency. Furthermore, this and other studies in which organ cultures were used (Tyrrell and Bynoe, 1966; Higgins, 1966; McIntosh et al., 1967; Higgins et al., 1970; Roome, Dickinson and Caul, 1971 ; Higgins and Ellis, 1972) were limited by the scarcity of human embryonic material and the laborious nature of the technique. The purpose of this paper is to report on the use of organ cultures in the study of a larger number of cases of acute infection of the respiratory tract. MATERIALS AND METHODS Specimens were collected and examined by standard met hods, as described previously (Higgins, Ellis and Boston, 1963; Higgins, Boston and Ellis, 1964). A nose and a throat swab were taken from each patient, placed in the same bottle of transport medium and conveyed to the laboratory on melting ice. The standard method of examination included inoculation onto a blood agar plate and into tissue cultures of monkey kidney, the Bristol line of HeLa cells, human-embryo diploid fibroblasts (Wl-38) and human-embryo kidney. The specimens were also inoculated subcutaneously and intracranially into suckling mice. Organ cultures of human-embryo nose and trachea were prepared initially as described by Hoorn but the method was later modified as recommended by Tyrrell and Blamire (1967). For the last 2 yr of the study all organ cultures were maintained in 6 x %-in. (15 x 1.5-cm) test tubes, instead of in 6-cm petri dishes, and transverse sections of trachea were used in preference to longitudinal sections (Higgins and Ellis). The fluids, harvested at intervals,

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