Human papillomavirus predicts outcome in oropharyngeal cancer in patients treated primarily with surgery or radiation therapy.

OBJECTIVE: This study examines the prognostic significance of human papillomavirus (HPV) in patients with locally advanced oropharyngeal squamous cell carcinoma (SCC) treated primarily with surgery or definitive radiotherapy. METHODS: One hundred and ninety-eight patients with Stage 3/4 SCC were followed up for recurrence in any form or death from any cause for between 1 and 235 months after diagnosis. HPV status was determined using HPV E6-targeted multiplex real-time PCR/p16 immunohistochemistry. Determinants of recurrence and mortality hazards were modelled using Cox's regression with censoring at follow-up dates. RESULTS: Forty-two per cent of cancers were HPV-positive (87% type 16). HPV predicted loco-regional control, event-free survival and overall survival in multivariable analysis. Within the surgery with adjuvant radiotherapy (n=110), definitive radiotherapy-alone (n=24) and definitive radiotherapy with chemotherapy (n=47) groups, patients with HPV-positive cancers were one-third or less as likely to have loco-regional recurrence, an event or to die of any cause as those with HPV-negative cancers after adjusting for age, gender, tumour grade, AJCC stage and primary site. The 14 patients treated with surgery alone were considered too few for multivariable analysis. CONCLUSION: HPV status predicts better outcome in oropharyngeal cancer treated with surgery plus adjuvant radiotherapy as well as with definitive radiation therapy±chemotherapy.

The diagnosis of chlamydia, gonorrhoea, and trichomonas infections by self obtained low vaginal swabs, in remote northern Australian clinical practice.

OBJECTIVE: To examine the diagnostic performance of self obtained low vaginal swabs (SOLVS) and polymerase chain reaction (PCR) techniques in the diagnosis of Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), and Trichomonas vaginalis (TV) infection in a variety of clinical practice settings in remote north western Australia. DESIGN: A cross sectional field study of microbiological collection techniques in women undergoing gynaecological investigation in remote settings performed by a variety of practitioner types over 10 months. PARTICIPANTS AND SETTING: 349 women from remote towns and communities in the Kimberley region of north west Western Australia having gynaecological examinations for clinical reasons, well women screening, antenatal screening, and sexual health examinations. RESULTS: The overall prevalence of infection in the study population based on any positive conventional sample was 9.2%, 7.6%, and 16.1% for CT, NG, and TV respectively. The detection rates for CT and NG by SOLVS were 89% and 96% respectively, compared with 79% and 91% for endocervical swabs and 79% and 83% for first void urine. SOLVS had a sensitivity of 93% for TV detection, equal to that of clinician obtained low vaginal swabs. None of these differences reached statistical significance. A combination of SOLVS and first void urine detected 96% of the CT cases, 100% of the NG cases, and 96% of TV cases. CONCLUSIONS: Self obtained low vaginal swabs are an acceptable, simple and sensitive diagnostic sample for the detection of CT, NG, and TV, and have particular applications in remote clinical practice and as a screening technique.

A large outbreak of conjunctivitis caused by a single genotype of Neisseria gonorrhoeae distinct from those causing genital tract infections.

Several epidemics of gonococcal conjunctivitis have occurred in Aboriginal populations in Central Australia. In 1997, the first outbreak in the Kimberley region of Western Australia occurred, spreading to Central Australia with a total of 447 cases. A genotyping method was applied directly to DNA extracted from patient samples to characterize the gonococcus causing the epidemic and to compare it with contemporaneous genital isolates. Those positive conjunctival specimens from Kimberley and Central Australia that could be genotyped were all indistinguishable, but were distinct from the genital gonococci, even when they shared the same auxotype and serotype. This suggested that the outbreak was due to a single genotype of Neisseria gonorrhoeae that had probably been carried between communities by infected individuals. We did not find evidence to support the existence of a genital reservoir of the types causing epidemic gonococcal conjunctivitis.

Diagnosis of Chlamydia trachomatis and Neisseria gonorrhoeae. Genitourinary infections in males by the Amplicor PCR assay of urine.

The Amplicor CT/NG polymerase chain reaction (PCR) test on urine specimens from males was prospectively evaluated against established specimens and laboratory methods for diagnosing Chlamydia trachomatis and Neisseria gonorrhoeae genitourinary infections, in patients from a remote region of Western Australia. Seventy-three males who were tested for both C. trachomatis and N. gonorrhoeae by both conventional methodology and Amplicor PCR on urine were enrolled in the study. Established testing comprised enzyme immunoassay/immunofluorescence antigen testing (EIA/IF) for C. trachomatis and microscopy and/or culture for N. gonorrhoeae on urethral swabs. Positive test results were confirmed using a set of criteria that included supplemental PCR testing and clinical history. Overall, 13.7% of patients were resolved as positive for C. trachomatis and 52.1% as positive for N. gonorrhoeae. The sensitivity and specificity of the Amplicor CT/NG PCR on male urine specimens for C. trachomatis were 80.0% (8/10) and 95.2% (60/63), compared with 60.0% (6/10) and 100.0% (63/63) for EIA/IF on urethral swabs. For N. gonorrhoeae, the sensitivity and specificity of the Amplicor CT/NG PCR on male urine specimens were both 100% (38/38 and 35/35, respectively) compared with 86.8% (33/38) and 100% (35/35) for microscopy and/or culture on urethral swabs. The results of this study indicate that the Amplicor CT/NG multiplex PCR test for C. trachomatis and N. gonorrhoeae performed on urine in males provides a highly sensitive, specific, and robust method for the diagnosis of both C. trachomatis and N. gonorrhoeae, for the early detection of both symptomatic and asymptomatic infected individuals.

The distribution of antibodies to HHV-6 compared with other herpesviruses in young children.

Sera from 141 infants aged 0-12 months were examined for IgG antibodies to HHV-6, HSV, CMV, VZV and EBV and for HHV-6 specific IgM. Following the decline in maternal antibody, antibody to HHV-6 was found to rise by 5-6 months and approached the level found in adults by 11-12 months. In contrast the antibody rates for the other herpesviruses were much slower to rise, especially in the case of CMV and EBV. HHV-6 IgM antibodies were detected mainly in age groups showing a rapid rise in antibody to HHV-6. HHV-6-IgM was not detected in 235 cord blood samples. The data suggest that HHV-6 infection is acquired horizontally, at a very early age in Western Australia.

Frequent shedding of human herpesvirus 6 in saliva.

We have previously reported the isolation of HHV-6 from saliva samples. Because these isolations were made in PHA-stimulated lymphocytes from healthy adults, which may occasionally contain endogenous HHV-6, it was desirable to repeat this work using cord blood lymphocytes. In this study 18 isolations of viruses provisionally characterized as HHV-6 were made from 19 saliva samples by centrifugally enhanced inoculation into PHA-stimulated cord blood lymphocytes. HHV-6 was not found in 10 pernasal aspirates, 50 endocervical swabs, or 30 male urethral swabs. It is concluded that HHV-6 is usually present in the saliva of most adults and that this affords a possible explanation of the high infection rate with this virus in young children.

Comparison of immunodiffusion and enzyme linked immunosorbent assay for antibodies to four Aspergillus species.

Antigenic extracts were prepared from Aspergillus fumigatus, A niger, A flavus and A terreus for use in enzyme linked immunosorbent assay (ELISA) and immunodiffusion (ID) tests for Aspergillus antibodies to determine whether the use of antigenic extracts from species other than A fumigatus increased the sensitivity of the ELISA. ELISA titres correlated well with positive ID tests. Patient titres by ELISA were significantly higher than control titres for all species. Patient titres to A niger were also significantly higher than titres to the other species. Total number of ID bands to A fumigatus correlated significantly with anti-A fumigatus ELISA titres. It is concluded that the use of antigenic extracts from species other than A fumigatus improves the sensitivity of the ELISA.

Efficient primary isolation of human immunodeficiency virus.

A highly-sensitive and efficient culture technique for human immunodeficiency virus type-1 (HIV-1) is described; HIV-1 was recovered from the lymphocytes of 44 (94%) antibody-seropositive healthy or symptomatic individuals. The reductions in the requirements for both the reagent volume and the number of patients' lymphocytes, together with an increased efficiency, has made this HIV-1 culture system more practical for diagnostic virology laboratories.

Centrifugal enhancement of human immunodeficiency virus (HIV) and human herpesvirus type 6 (HHV-6) infection in vitro.

The effect of centrifugal inoculation of human immunodeficiency virus (HIV) and human herpesvirus-6 (HHV-6) on the infectivity of the viruses for cell cultures was examined. Three HIV-1 strains, ARV-2, HTLV-IIIb and a local isolate, WA-46c, were tested in peripheral blood lymphocytes, HUT-78, H9 and MT-2 cells. The HHV-6 strain was a local isolate and was studied only in peripheral blood lymphocyte cultures. Centrifugal inoculation of the viruses at a force of 2500 x g for 60 min, enhanced HIV-1 infectivity by a factor of about 10-fold in all cell cultures tested. Infectivity was increased about 100-fold for HHV-6.

Human herpes virus type 6 (HHV-6) and its in vitro effect on human immunodeficiency virus (HIV).

Human herpes virus type 6 (HHV-6) was isolated from the peripheral blood lymphocytes of a patient infected with human immunodeficiency virus (HIV). Antibodies to this herpes virus were found to be widespread among adults and children in Western Australia. Co-infection studies indicated that HIV replication was inhibited by the presence of HHV-6.

Importance of virus passage history on the growth of Coxsackie B4 in human skin fibroblasts.

Human skin fibroblasts have previously been reported to display an age-dependent resistance to infection with coxsackie B4 virus. We have shown that the virus will replicate and produce CPE in human skin fibroblasts regardless of the age of the donor of the cells. The passage history of the virus was found to influence the titre of the virus in these cells.

A modified immunofluorescence test for Epstein-Barr virus-specific IgM antibody.

The fluorescent antibody (FA) test for Epstein-Barr virus (EBV)-specific IgM antibody was improved by the use of sodium butyrate to induce a higher level of EBV antigen expression in P3HR-1 slide preparations and by removal of rheumatoid factor (RF) and IgG antibodies from test sera by means of adsorption with suspensions of Sepharose-IgG and Streptococcus pyogenes strain AR1. This method was compared with the Paul-Bunnell test (PB) on 1106 sera submitted to a routine virus diagnostic laboratory for infectious mononucleosis serology and 96.4% of sera showed concordant results. Thus the EBV-IgM-FA method was suitable for routine diagnostic use. However, it proved helpful to test EBV-IgM positive sera by PB to assist in the detection of cross-reacting IgM antibodies sometimes present.

Electron microscopy after direct ultracentrifugation.

A direct ultracentrifugation technique was used in the preparation of skin lesion specimens for examination by electron microscopy. The concentration factor of centrifuged specimens was estimated to be in excess of 1,000-fold compared to conventional adsorption techniques. This resulted in an increase of over 300% in the detection rate of herpesviruses and poxviruses from skin lesion specimens.

Association of genital adenovirus infection with urethritis in men.

Adenoviruses were isolated from the urethral swabs of 129 male patients in an STD clinic. After exclusion of patients with Chlamydia trachomatis or Neisseria gonorrhoea infections, 85 of the remaining 120 patients had urethritis, compared with 28 men with urethritis detected in a control group which was closely matched for age, sex, and date of specimen collection. This statistically significant difference suggests that genital adenovirus infection may be a cause of urethritis in some male patients.

False positive results occurring in a radioimmunoassay for hepatitis A IgM antibody of the IgM class.

The diagnosis of hepatitis A infection is usually based on the presence of hepatitis A specific IgM in a single serum sample. The fortuitous observation in one patient that this reactivity was apparently still present 19 mth after her original illness led to the discovery that the ABBOTT HAVAB-M kit method may produce false positive results. A series of patients who had previously had hepatitis A was retested and false positive results were found in 6% of this group. Control groups consisted of patients with other acute and chronic liver disorders and other acute viral diseases. No reactivity was detected in the control sera. Sucrose gradient fractionation revealed that the factor responsible for the false positive results was associated only with serum fractions containing IgA and IgG and that it could be removed by absorption of sera with staphylococcal protein A but not by absorption with streptococcus AR1 or by 2-mercaptoethanol treatment. It was concluded that following hepatitis A infection some patients produce a rheumatoid factor-like substance (not of IgM class) which is cleared from the serum in 2-3 yr. The presence of this factor may lead to a misdiagnosis in patients presenting with jaundice.

Variation in murine cytomegalovirus replication in fibroblasts from different mouse strains in vitro: correlation with in vivo resistance.

The in vitro growth of murine cytomegalovirus (MCMV) in mouse embryo fibroblasts (MEF) from Balb/c, C57BL, C3H and CBA mouse strains has been shown to correlate with the genetically determined in vivo susceptibility of these strains to MCMV infection. MEF from Balb/c and C57BL mice were more susceptible to MCMV than those from C3H and CBA mice. This was evident regardless of whether replication was measured by cytopathic effect (c.p.e.) score, virus yield, plaque count, plaque size or time of onset of c.p.e. The growth of MCMV in tracheal organ cultures from different mouse strains was similar to that observed in MEF from these strains. The replication in MEF when measured by c.p.e. score and virus yield was affected by the density of the cell cultures. The strain of mouse used to produce MCMV also affected the comparative sensitivity of MEF to the virus. This appeared to be due to reduced growth of MCMV in its homologous MEF type, an unexpected result. In contrast, cell density had little effect on the replication of herpes simplex virus type 1 (HSV-1), but again the in vitro susceptibility of MEF reflected the in vivo susceptibility of mouse strains to infection with this virus.

Acute haemorrhagic cystitis caused by adenovirus type 11 in a recipient of a transplanted kidney.

A case of haemorrhagic cystitis caused by adenovirus type 11, which occurred in a female patient 13 weeks after renal transplantation is described. The patient lacked neutralising antibody to adenovirus type 11 before transplantation, but this antibody was present in the serum of the kidney donor. The patient developed antibody to adenovirus type 11, and this virus was isolated from her throat and urine. The data suggest that the infection was transmitted by the kidney graft.