Sinusitis in the common cold.

BACKGROUND: Acute community-acquired sinusitis is considered a bacterial complication of the common cold. Radiologic abnormalities in sinuses occur, however, in most patients with upper respiratory virus infections. OBJECTIVE: Assessment of the occurrence, clinical profile, laboratory findings, and outcome of radiologically confirmed sinusitis was carried out as part of a common cold study in young adults. METHODS: Clinical examinations and radiography of the paranasal sinuses were carried out on days 1, 7, and 21 in 197 patients with the common cold. The symptoms were recorded on diary cards on days 1 to 20. Ten viruses and 5 bacteria were studied as etiologic agents of common cold as reported earlier. Serum C reactive protein concentrations, erythrocyte sedimentation rates, and total white blood cell counts with differentials were determined in 40 randomized subjects on day 7. The effect of 6 days of intranasal fluticasone propionate treatment of the common cold in the prevention of sinusitis was analyzed. RESULTS: On day 7, 39% of patients with the common cold in the placebo group (n = 98) had sinusitis, which we would prefer to call viral sinusitis. The symptoms of patients with sinusitis and those without it were not clinically distinguishable. Viral infection was detected in 81.6% of patients with sinusitis. No significantly increased levels of antibodies to bacteria were detected. Serum C reactive protein concentrations, erythrocyte sedimentation rates, and white blood cell counts were low in patients with sinusitis. All patients made a clinical recovery within 21 days without antibiotic treatment. Fluticasone propionate treatment tended to prevent paranasal sinusitis, especially in rhinovirus-positive subjects. CONCLUSION: Viral sinusitis frequently occurs in the early days of the common cold, but it is a self-limited illness. The sinuses should not be imaged in patients with the common cold if the signs and symptoms of illness gradually become less severe and no specific signs suggestive of bacterial sinusitis occur.

Molecular diagnosis of human rhinovirus infections: comparison with virus isolation.

To compare the sensitivity and specificity of RT-PCR with that of virus isolation in the detection of human rhinoviruses, we tested nasopharyngeal aspirates from 200 patients on the 1st and 7th days after the onset of the common cold. An assay utilizing a short amplicon in the conserved 5' noncoding region was found highly sensitive. Of 192 positive samples altogether, 65 were found positive by RT-PCR only, 6 were positive by isolation exclusively, and 121 gave positive results in both tests.

Viruses and bacteria in the etiology of the common cold.

Two hundred young adults with common colds were studied during a 10-month period. Virus culture, antigen detection, PCR, and serology with paired samples were used to identify the infection. Viral etiology was established for 138 of the 200 patients (69%). Rhinoviruses were detected in 105 patients, coronavirus OC43 or 229E infection was detected in 17, influenza A or B virus was detected in 12, and single infections with parainfluenza virus, respiratory syncytial virus, adenovirus, and enterovirus were found in 14 patients. Evidence for bacterial infection was found in seven patients. Four patients had a rise in antibodies against Chlamydia pneumoniae, one had a rise in antibodies against Haemophilus influenzae, one had a rise in antibodies against Streptococcus pneumoniae, and one had immunoglobulin M antibodies against Mycoplasma pneumoniae. The results show that although approximately 50% of episodes of the common cold were caused by rhinoviruses, the etiology can vary depending on the epidemiological situation with regard to circulating viruses. Bacterial infections were rare, supporting the concept that the common cold is almost exclusively a viral disease.

Waterborne outbreak of viral gastroenteritis.

A waterborne epidemic took place in a Finnish municipality in April 1994. Some 1500-3000 people, i.e. 25-50% of the population, had symptomatic acute gastroenteritis. Laboratory findings confirmed adenovirus, a Norwalk-like agent, small round viruses (SRV), and group A and C rotaviruses as causative agents, Norwalk virus being the main cause of the outbreak. The epidemic was most probably associated with contaminated drinking water. The groundwater well, situated in the embankment of a river, was contaminated by polluted river water during the spring flood. A back flow from the river to the well had occurred via a forgotten drainage pipe.

Clinical scoring system in the evaluation of adult pharyngitis.

OBJECTIVE: To compare results of a clinical scoring system for diagnosis of group A streptococcal pharyngitis with microbiologic results, when several different pharyngeal pathogens were tested simultaneously. DESIGN: Evaluation of clinical manifestations of 106 adult patients with pharyngitis of different microbial origin. SETTING: General private practice; Health Center Pulssi, Turku, Finland. PATIENTS: Adult patients whose chief complaints were sore throats. MAIN OUTCOME MEASURE: A symptom score that was assigned to each patient according to the total number of certain signs and symptoms that are postulated to increase the probability of group A streptococcal pharyngitis and blood measurements for infection. RESULTS: The highest symptom scores, 3 and 4, were found in 21 patients. These patients had pharyngitis due to group A streptococcus (four patients), group C streptococcus (four patients), group G streptococcus (two patients), group F streptococcus, Mycoplasma pneumoniae, Chlamydia pneumoniae, influenza A virus, influenza B virus, herpes simplex type 1 virus (two patients), and coxsackie B4 virus. No pathogen could be identified from three of the 21 patients. The C-reactive protein values and the leukocyte counts were raised significantly more often in streptococcal infections than in infections of other origin; the P values were .00016 and .028, respectively. CONCLUSION: Use of a clinical scoring system alone for diagnosis of pharyngitis may lead to improper use of anti-microbial agents. There is a need for accurate microbiologic diagnostic procedures in general practice to determine proper treatment of pharyngitis as well as to test the effect of antibacterial and, in the future, antiviral treatment in respiratory tract infections.

Expression of the L2 and E7 genes of the human papillomavirus type 16 in female genital dysplasias.

The expression of the E7 and L2 genes of HPV 16 was studied in benign and precancerous female genital lesions to evaluate their role in the development of dysplasias. Ninety biopsy specimens from 70 patients, selected on basis of dot blot DNA hybridization, were included in immunohistochemical and in situ hybridization analyses. In the HPV 16 DNA positive cases, L2 mRNA and E7 mRNA were detected in biopsies from 24 and 21 patients, respectively. L2 mRNA was found in eight of 16 cases of condyloma and mild dysplasia, and in 13 of 14 cases of moderate to severe dysplasia. The figures for E7 mRNA were 6/16 and 13/14, respectively. We found L2 mRNA in four of 12 normal or condylomatous specimens and E7 mRNA in only one of these. The detection rates for L2 and E7 mRNAs increased along with the severity of the lesions (P = 0.0064 and P = 0.0001, respectively). The L2 protein was found in one condyloma and in 12 dysplasias, eight of which were moderate or severe. The L2-antibody-reactive cells were localized in superficial layers of the epithelium. The detection rate for L2 mRNA and especially for E7 mRNA increased along with the histopathologic grade of the lesion.

Surveillance of human immunodeficiency virus (HIV) antibodies in medicolegal autopsies in Finland--monitoring early changes in HIV-seropositivity among risk groups and average population.

In order to cooperate with voluntary screening programs aimed at the surveillance of the HIV epidemic in Finland, we have studied medicolegal autopsies for HIV antibodies since 1986 using an enzyme immunoassay on postmortem sera. The investigation covered 47.4% and 39.2%, respectively, of all deaths under the age of 65 years in the metropolitan areas of Helsinki and Turku--two cities on the densely populated southern coast of Finland from which most HIV infections have thus far been detected. Nine HIV-positive cases (0.12%) were detected among the 7305 medicolegal autopsies tested in 1986 to 1990. This figure is higher than the prevalence of 0.01 to 0.03% in voluntary screening programs for the general population would suggest. Seven of our cases had previously tested positive, and two were previously unknown cases, indicating that people at high risk are clustered in the medicolegal autopsy series. Of the six cases in an early stage of infection, three committed suicide suggesting the importance of HIV-screening in suicide cases in tracing symptomless HIV carriers. Five of the cases were detected in 1990, a year when the number of new HIV infections had more than doubled compared to the previous two years. This suggests that testing of medicolegal autopsies as surrogate tests for the population gives useful information even in low-prevalence areas like Finland. Such testing has none of the ethical problems of many other back-up surveys, and may be particularly sensitive to early changes in epidemiology.

Human papillomavirus detection from the female genital tract: combined vs. separate scrape methods.

Because genital human papillomavirus (HPV) infections tend to be multifocal, it was studied how effective one combined specimen is in detecting HPV-DNA from the lower female genital tract. The study population consisted of 50 patients referred to a colposcopy clinic for a suspected condylomatous and/or dysplastic lesion. From half of the patients, a separate scrape from the cervix, vagina and vulva was taken first followed by a combined scrape representing all the genital sites, and from the other half, vice versa. HPV-DNA (types 6, 11, 16 and 18) was identified using the AffiProbe hybridisation test. Thirty-six specimens collected from 17 patients were positive for HPV-DNA. A multifocal infection was demonstrated in at least 11/17 (65%) HPV-DNA positive patients. The combined scrape was the most informative specimen, revealing 75% of all HPV-DNA-positive patients. It was concluded that HPV-DNA can reliably be detected from the female genital tract in a simple way from one combined specimen.

Detection of human papilloma virus (HPV) DNA in genital biopsy specimens by in situ hybridization with digoxigenin-labeled probes.

The presence of human papillomavirus (HPV) nucleotide sequences in paraffin sections of genital biopsies was examined by in situ hybridization using non-isotopic, digoxigenin-labeled probes representing HPV types 11, 16 and 18. Digoxigenin-labeling of the probes was performed using DNA labeling and a commercially provided detection kit. Hybridization was performed under stringent conditions. The hybrids were detected by using anti-digoxigenin alkaline phosphatase conjugate and visualized with enzyme catalyzed color reaction. In situ hybridization with digoxigenin-labeled probes was a useful technique for identification of HPV infection. The results were compared with the results obtained with radiolabeled DNA probes. The sensitivity of the digoxigenin-labeled probes was equal to the sensitivity of the radiolabeled probes. The background with digoxigenin-labeled probes was very low. Using nonradioactive probes the localization of hybrids at the cellular level was better than 35S-labeled probes.

Polyethylene glycol increases specificity of hybridization typing of HPV specimens.

Different hybridization conditions for the typing of human papillomaviruses (HPV) in clinical biopsy specimens were tested. The stringency of the hybridization reaction was varied by altering the formamide concentration in the solution. Two polymers, dextran sulphate and polyethylene glycol (PEG), were compared as accelerators of the hybridization reaction. The PEG-containing hybridization solution was found to be suitable for typing clinical HPV specimens. Single-stranded RNA probes proved to be more specific than DNA probes in typing clinical specimens. Specimens that were positive both for HPV6 and HPV11 in spot hybridization were confirmed by Southern hybridization. In total, 467 biopsy specimens from genital, anal, oral, aural and nasal lesions were examined for HPV6, HPV11, HPV16 and HPV18 DNA by spot hybridization. Approximately one-third of the specimens were positive for these types.

Pharyngitis in adults: the presence and coexistence of viruses and bacterial organisms.

STUDY OBJECTIVE: To determine the presence and coexistence of viruses and bacterial organisms causing pharyngitis in adults. DESIGN: Open study using diagnostic methods, including rapid antigen-detection techniques, to test for the presence of viruses of the respiratory tract, as well as Mycoplasma pneumoniae. Chlamydia trachomatis, the Chlamydia species strain TWAR, and beta-hemolytic streptococci. SETTING: Open health care. PATIENTS: One hundred six consecutive adult patients, 15 to 65 years old, whose chief complaint was sore throat. MAIN RESULTS: Of the 106 patients, beta-hemolytic streptococci were found in only 24 patients (5 patients with group A streptococci, 13 with group C, 5 with group G, and 1 with group F); M. pneumoniae was found in 10 patients, the Chlamydia species strain TWAR in 9 patients, and viruses in 27 patients. Two microbes were simultaneously isolated in 3 patients, and no microbial findings were detected in 33 patients. CONCLUSION: Because 19 patients were infected with the Chlamydia species strain TWAR and M. pneumoniae, and 24 patients were infected with beta-hemolytic streptococci, the diagnostic procedures and therapies for adult patients with pharyngitis need to be reconsidered. The results of our study also confirm earlier suggestions that the Chlamydia species strain TWAR alone is a causative agent for pharyngitis in adults.

Comparison of smear specimens with biopsy specimens in a nucleic acid hybridization test for human papilloma virus (HPV) infection.

A total of 323 pairs of specimens from women with Papanicolaou class II or III cytology were examined for human papillomavirus (HPV) types 6, 11, 16 and 18 by spot hybridization. Each pair consisted of a representative biopsy specimen and a smear specimen from cervical, vaginal or, more rarely, vulvar lesions. We found a close correlation between HPV findings in biopsies and smears. In 83.9% (271) of the cases, both specimens were either positive for a given HPV type or negative. No discordant HPV-types in the two types of specimens were found. In 15.8% (51) of the cases, one specimen proved positive for a given HPV-type while the other specimen from the same patient was negative. In 8.0% (26) of the cases, the biopsy specimen proved positive and the corresponding smear specimen was negative. On the other hand, in 7.7% (25) of the cases we were able to detect HPV DNA in the smear specimen, whereas the corresponding biopsy specimen was negative. We suggest that a smear specimen would be advantageous for screening large groups of patients for the presence of a HPV infection in the genital tract. By using smear specimens together with biopsy specimens it is possible to maximize the number of HPV infection diagnoses.

Herpes simplex virus detection by macroscopic reading after overnight incubation and immunoperoxidase staining.

Human diploid foreskin fibroblast cells grown in 24-well plates were inoculated with clinical specimens by centrifugation at 1,000 X g for 45 min. Cultures were incubated at 37 degrees C overnight, fixed, and stained with peroxidase-labeled monoclonal antibodies against herpes simplex virus types 1 and 2. Stained plaques of infected cells were large enough to be detected with the naked eye, and microscopic examination did not reveal any further positive specimens. The method was compared with standard isolation in human fibroblasts grown in shell vials and inoculated by centrifugation at 4,000 X g, observed microscopically for the occurrence of typical cytopathogenic effect three times a week for 10 days, and then typed by enzyme immunoassay. Of the 289 specimens tested, 105 were positive and 174 were negative by both methods. Six specimens were positive by standard isolation only, two of them containing varicella-zoster virus, and two specimens were stored frozen before being tested by immunoperoxidase staining. Two specimens found negative by standard isolation were positive by immunoperoxidase staining. For two specimens negative by immunoperoxidase staining, the standard isolation cultures were lost due to microbial contamination. Forty-two specimens found positive by standard isolation were clearly positive when stained only 8 h after inoculation. By standard isolation, positive results were reported on the average 3 to 4 days after inoculation, whereas by immunoperoxidase staining the result was available within less than 24 h. Immunoperoxidase staining of infected cells is a sensitive method for rapid laboratory diagnosis of herpes simplex virus infections, and 24-well plates are convenient for the handling of a large number of specimens.

Characterization of the common immunodominant antigenic determinant in the lipopolysaccharide of Bacteroides fragilis by a monoclonal antibody.

The target determinant of a monoclonal antibody (MoAb) to Bacteroides fragilis lipopolysaccharide (LPS) was characterized by inhibition enzyme immunoassay (EIA), immunoblotting (IB), immunofluorescence technique (IF) and electron immunocytochemical (EIC) technique. The MoAb has been shown to react positively with 96% of B. fragilis isolates. LPS preparations from 14 different B. fragilis strains were tested by EIA and IB. Two LPS preparations did not react in any of the tests. In both preparations the D-galactose was either lacking or present in low amount compared with the other LPSs. In addition, inhibition experiments with synthetic disaccharides confirmed that the target determinant is composed of beta-1,6-linked galactose disaccharide. EIC showed that the target of the LPS-MoAb is located on the surface of the outer membrane. These results show that the galactose chain present in LPS isolated from most B. fragilis strains contains the immunodominant antigenic determinant.

Comparison of polyclonal and monoclonal antibodies to Actinomyces and Arachnia species.

Polyclonal (PoAbs) and monoclonal (MoAbs) antibodies were produced to Actinomyces israelii serotypes 1 and 2, to Actinomyces naeslundii, and to Arachnia propionica, and their specificities were studied by an enzyme immunoassay (EIA). All PoAbs except those to A. propionica reacted also with at least one other Actinomyces species. Only the MoAb to A. naeslundii proved to be more specific than the corresponding PoAbs. This MoAb did not crossreact with other Actinomyces or Arachnia species, nor with any other anaerobic or aerobic bacteria studied by inhibition EIA. Immunoblotting studies indicated that the antibody specific to A. naeslundii is directed against a large molecular weight antigen (greater than 150 kd), probably polysaccharide in nature. The produced PoAbs and MoAbs can be used for further analyses of the antigenic determinants of different Actinomyces and Arachnia species.

Typing of herpes simplex virus isolates with monoclonal antibodies and by nucleic acid spot hybridization.

Fifty-one clinical isolates of herpes simplex virus (HSV) were typed by an enzyme immunoassay (EIA) using mouse monoclonal antibodies, by DNA spot hybridization, and by restriction enzyme analysis using restriction endonuclease Eco RI. Extracts of VERO cells infected with the isolates were used for coating microtitre plates or denatured and spotted onto nitrocellulose filters. Viral antigens passively adsorbed to microtitre plates were detected by an indirect EIA using mouse monoclonal antibodies specific for HSV type 1 (HSV-1) or HSV type 2 (HSV-2). Spotted DNA was hybridized with 32P-labeled probes containing Hind III/Sal I-fragments of either HSV-1 or HSV-2 DNA and bound radioactivity was detected by autoradiography and counted in a liquid scintillation counter. All the three methods gave identical results for the 51 isolates studied. Twenty-six isolates were identified as HSV-1 and 25 as HSV-2. An additional 30 specimens were tested only by EIA and hybridization. Results by both techniques were in complete agreement.

Monoclonal antibodies to Bacteroides fragilis lipopolysaccharide.

Monoclonal antibodies (MoAbs) to the lipopolysaccharide (LPS) of Bacteroides fragilis were produced by immunizing mice before hybridization with bacterial outer membranes solubilized with Triton X-100. Nineteen stabile clones were established. They all produced antibodies that reacted more strongly with purified B. fragilis LPS than with crude sonicated antigen in an enzyme immunoassay. Four MoAbs were studied by immunoblotting and enzyme immunoassay inhibition. Immunoblotting confirmed that the target of the MoAbs was LPS. Marked and homogeneous staining occurred in the immunoblotting with both purified LPS and outer membranes in the molecular weight range of 8,000 to 27,000. In enzyme immunoassay inhibition, MoAbs reacted positively with 93 to 96% of B. fragilis strains, including prototype strains ATCC 23745 and NCTC 9343. Within the B. fragilis group, the MoAbs reacted positively with two of five B. ovatus strains and two to six of nine B. thetaiotaomicron strains. No marked cross-reactivity with other bacteria was observed. These results confirm earlier findings that the B. fragilis LPS contains an immunodominant antigenic determinant common to almost all B. fragilis isolates.

Viral diagnoses using the rapid immunofluorescence technique and epidemiological implications of acute respiratory infections among children in different European countries.

From November 1978 to October 1981, a total of 7716 specimens of nasopharyngeal secretions were examined by the rapid immunofluorescence technique to determine the frequency of infections caused by the respiratory syncytial virus (RSV), influenza virus A, and parainfluenza viruses 1 and 3. The tests were carried out in six different virus laboratories located in Newcastle upon Tyne (England), Copenhagen, Oslo, Stockholm, Turku (Finland), and Vienna; laboratories in Lisbon and Paris participated in the study for shorter periods. The specimens were collected from infants and children less than 6 years of age who had been admitted to hospital with an acute respiratory infection. Standardized techniques and quality controlled reagents were used. At least one of the above viruses was detected in 1927 (25%) of the specimens: RSV in 1475, influenza virus A in 123, parainfluenza virus 1 in 110, and parainfluenza virus 3 in 237 specimens. Respiratory syncytial virus dominated in all centres, but in some Scandinavian centres distinct outbreaks due to this virus occurred only once or twice during the 3 years' study period. Three outbreaks of RSV were observed in Newcastle, but here an unprecedented delay of the first winter's epidemic occurred. The delay was associated with prolonged school closures in the area, and with a very early outbreak of influenza. Parainfluenza virus 3, which was predominantly a summer virus in Newcastle, was most frequently encountered during the colder months of the year in the other centres.