A new confirmatory Neisseria gonorrhoeae real-time PCR assay targeting the porA pseudogene.

The Roche Cobas Amplicor system is widely used for the detection of Neisseria gonorrhoeae but is known to cross react with some commensal Neisseria spp. Therefore, a confirmatory test is required. The most common target for confirmatory tests is the cppB gene of N. gonorrhoeae. However, the cppB gene is also present in other Neisseria spp. and is absent in some N. gonorrhoeae isolates. As a result, laboratories targeting this gene run the risk of obtaining both false-positive and false-negative results. In the study presented here, a newly developed N. gonorrhoeae LightCycler assay (NGpapLC) targeting the N. gonorrhoeae porA pseudogene was tested. The NGpapLC assay was used to test 282 clinical samples, and the results were compared to those obtained using a testing algorithm combining the Cobas Amplicor System (Roche Diagnostics, Sydney, Australia) and an in-house LightCycler assay targeting the cppB gene (cppB-LC). In addition, the specificity of the NGpapLC assay was investigated by testing a broad panel of bacteria including isolates of several Neisseria spp. The NGpapLC assay proved to have comparable clinical sensitivity to the cppB-LC assay. In addition, testing of the bacterial panel showed the NGpapLC assay to be highly specific for N. gonorrhoeae DNA. The results of this study show the NGpapLC assay is a suitable alternative to the cppB-LC assay for confirmation of N. gonorrhoeae-positive results obtained with Cobas Amplicor.

[Transport of newborn infants who need intensive care]. / Intenzív ellátást igényló újszülöttek szállítása.

A 4-year-long experience is reported on mobile neonatal emergency transport service for pathological newborn babies from referral hospitals to the NICU level III. The area covered by the NICU consists of three counties with a total population of 1 million. Distance of the hospitals from the NICU ranges 0.5-115 km, transport time varies from 0.5-5.0 hours. In the transport staff are dedicated neonatal team from the NICU III, the driver and the emergency medical technician are the employee of the National Ambulance Service. During the study period 385 patients were transported. Clinical condition and characteristic laboratory parameters of the babies at departure and arrival are analysed and compared to evaluate the efficacy and quality of the system. Furthermore, some questions regarding organisation, transit time and response time and legal aspects are discussed.

Further observations on the immune response to recombinant hepatitis B vaccine after administration to aboriginal and Torres Strait Island children.

OBJECTIVE: To determine the prevalence of markers of hepatitis B virus (HBV) immunity and infection at 5 years of age in Aboriginal and Torres Strait Island children who were fully vaccinated in infancy, and to examine the response to a booster dose of hepatitis B vaccine in those children who had no detectable immunity despite vaccination. METHODOLOGY: A cross-sectional study of serological markers to HBV in a sample of 239 Aboriginal and Torres Strait Island children, with a mean age of 5.7 years, who were fully vaccinated in infancy. The antibody response to a booster dose of hepatitis B vaccine was determined in those children in the sample who had no markers of either immunity to HBV or infection with HBV. RESULTS: Of the 239 children, 6% (95% CI 4-10%) had been infected and, of these, four were HBV surface antigen (HBsAg) positive. Of the remaining 224 children, only 41% (95% CI 35-48%) had evidence of immunity (i.e. an antibody to HBV surface antigen (anti-HBs) level of > or = 10 miu/mL) to HBV. Of the children with no detectable immunity (i.e. anti-HBs < 10 miu/mL), 113 were followed up after receiving a booster dose of hepatitis B vaccine. Of these, 84% (95% CI 76-90%) had an anamnestic response (i.e. anti-HBs < 10 miu/mL following the booster dose). Therefore 16% (95% CI 10-24%) still had no detectable immunity following the booster dose. CONCLUSIONS: This study provides further evidence that Aboriginal and Torres Strait Island children have a suboptimal response to recombinant hepatitis B vaccine. It also indicates that a considerable number of Aboriginal and Torres Strait Island children in the study age cohort have been exposed to HBV. However, despite these concerns, this study and historical data provide strong evidence that there has been a marked reduction in the prevalence of HBV infection and carriage in previously 'high risk' Aboriginal and Torres Strait Island children since the introduction of hepatitis B vaccines. Aboriginal and Torres Strait Island children who have been fully vaccinated in infancy do not require a booster dose of hepatitis B vaccine at school entry.

Assessment of the RSCL quality of life instrument during chemotherapy in an Italian setting.

Evaluation of the quality of life of oncological patients is now perceived as a major element of cancer treatment, and various instruments for measuring it have been proposed. However, whatever instrument is used must be capable of reflecting the different ways in which different contexts and cultures respond to cancer. We administered the Italian version of the Rotterdam Symptom Checklist (RSCL) to 60 patients with advanced disease attending five oncological centres in Romagna (central Italy). The RSCL was found to be able to measure both psychological and physical well-being and to detect a difference between patients on low/medium and those on high-burden chemotherapy. The Italian version of the RSCL is an adequate scale that is capable of capturing patient perceptions and could be used for monitoring the quality of life of Italian patients participating in clinical trials.

Comparison of four methods for quantitative measurement of hepatitis B viral DNA.

AIMS/METHODS: Four assays for measuring HBV-DNA quantitatively have been compared with regard to sensitivity, precision and linearity. The methods were 125I-labelled solution hybridisation assay (liquid hybridisation, Abbott), an ELISA-based chemiluminescent RNA-DNA hybrid assay (RNA-DNA, Digene), a chemiluminescent branching oligonucleotide assay (bDNA, Chiron) and a membrane hybridisation assay using slot-blot equipment (slot blot). RESULTS: The bDNA assay was linear over three orders of magnitude and was the most sensitive assay, being approximately ten times more sensitive than the other assays, so that samples negative on RNA-DNA, liquid hybridisation and slot blot gave quantifiable results on bDNA. Furthermore, intra- and inter-assay variability showed that the bDNA and liquid hybridisation assays had the greatest precision, with coefficients of variation of 6.6% to 11.5% and 2.3% to 10.5%, respectively. However, the nominated amounts of HBV DNA in the standards (from all assays) were not reproducible in the other assays, such that amounts measured with bDNA would give values approximately twice that of RNA-DNA and 60 times that of liquid hybridisation. CONCLUSIONS: The recently developed bDNA assay has advantages compared with the other assays in quantitating samples with low levels of virus present. In addition, since the assays vary considerably by a number of criteria, the method of measurement should always be reported.

An ELISA for the detection of antibody to the core antigen of hepatitis C virus: use in diagnosis and analysis of indeterminate samples.

AIMS: In order to examine more carefully the natural history of hepatitis C virus infection and to determine a role for anti-core in the discrimination of indeterminate samples, a solid phase ELISA to detect antibody of the immunoglobulin G class to the hepatitis C virus core antigen was developed using purified protein expressed in Escherichia coli from a major portion of the core antigen coding region. METHODS/RESULTS: In a study which examined 651 samples submitted for routine testing by a commercial ELISA (Ortho), only 11 samples showed discrepant results; of these, 10 were Ortho ELISA positive, anti-core negative and one was Ortho ELISA negative anticore positive. Supplemental tests showed that 5/10 of these samples were anti-HCV negative by RIBA and the reciprocal 5 were negative for anti-C22 but positive for anti-C100 and anti-C33. The Ortho ELISA negative, anticore positive sample was weakly positive for anti-C22. The anti-core ELISA was then used to examine 67 indeterminate samples from the blood bank; 11/11 samples which were HCV-RNA positive were anti-core positive and 7/56 samples which were HCV-RNA negative were anti-core positive. The anti-core titre was then examined in two groups of indeterminate samples; group 1, polymerase chain reaction-positive, anti-core positive and group 2, polymerase chain reaction-negative, anti-core positive. The geometric mean anti-core titres in these groups were 1 x 10(-3.6) and 1 x 10(-2.3), respectively. Thus in this group of indeterminate samples, all samples (except one) with an anti-core titre > or = 1/200 were polymerase chain reaction-positive, confirming a close correlation between anti-core levels and hepatitis C viraemia. Anti-core was detected with equal efficiency in patients infected with genotypes which differed to that used to express the recombinant core antigen.

Immunity to hepatitis B, poliomyelitis and measles in fully vaccinated aboriginal and Torres Strait Island children.

OBJECTIVE: To determine the immunity to hepatitis B, poliomyelitis and measles in fully vaccinated Aboriginal and Torres Strait Island children in north Queensland. METHODOLOGY: A cross-sectional survey of immunity in a sample of children; 101 fully vaccinated Aboriginal and Torres Strait Island children, with a median age of 24.5 months, from 10 communities in North Queensland participated in this study. The main outcome measures were the prevalence of adequate antibody levels against hepatitis B, poliomyelitis and measles. RESULTS: Only 54% (95% CI 44-63%) of the children had adequate immunity (> or = 10 m iu/mL) to hepatitis B, and one child had been infected despite vaccination. Although all the children (95% CI 96-100%) had adequate immunity (i.e. neutralizing antibodies at a dilution of > or = 1:8) to poliovirus 2, only 93% (95% CI 86-96%) and 60% (95% CI 50-69%) had adequate immunity to polioviruses 1 and 3, respectively. Nearly all (96%; 95% CI 90-98%) of the children had adequate immunity (i.e. detectable IgG antibody) to measles. CONCLUSIONS: Although a relatively low proportion of the children had adequate antibody levels against hepatitis B the clinical significance of this observation is uncertain. Further studies are needed to determine whether fully vaccinated Torres Strait Island children have been adequately protected and whether they require a booster dose of hepatitis B vaccine. A substantial proportion of fully vaccinated Aboriginal and Torres Strait Island children are inadequately protected against poliomyelitis, and therefore any such child with acute flaccid paralysis should be investigated fully for poliomyelitis. Vaccinated Aboriginal and Torres Strait Island children are well protected against measles, as are other Australian children.