Xpert Xpress Flu/RSV: Validation and impact evaluation at a large UK hospital trust.

Despite vaccination programs and antivirals, influenza remains a prominent cause of morbidity and mortality. The Xpert Xpress Flu/respiratory syncytial virus (RSV) test is a leading influenza point-of-care test, but its evaluation has been limited to nasopharyngeal samples. In addition, the clinical impacts of Xpress Flu/RSV have not yet been quantified. We evaluated the performance of Xpress Flu/RSV at three locations in a UK Hospital Trust against an existing laboratory assay. Multiple upper respiratory tract sample types were included. In addition, we calculated time saved by Xpert, and the associations between Xpert use and rates of early patient isolation and antiviral prescription as recorded at the time of the laboratory result being telephoned out. A total of 642 patients were included in the diagnostic performance analysis. There were 177 laboratory-confirmed cases of influenza A, 7 influenza B and 86 RSV. For influenza A, sensitivity and specificity were 96.6% (95% confidence interval [CI]: 92.8%-98.8%) and 98.1% (CI: 96.4%-99.1%), respectively. This was sustained across all locations and sample types. The negative predictive value was 98.7% (CI: 97.2%-99.4%). The median amount of time saved was 27.1 h. Xpert use was associated with sixfold higher rates of isolation and threefold higher rates of antiviral prescribing by the time the laboratory result was available. Sensitivity for RSV was lower at 86.0% (95% CI: 76.9%-92.6%). Xpert Xpress Flu/RSV reliably detects influenza A infection and has significant clinical impacts. Cartridge optimization is required to enable accurate multiplexing, including from a range of sample types.

Evaluation of Vela Diagnostics HIV-1 genotyping assay on an automated next generation sequencing platform.

BACKGROUND: Antiretroviral drug resistance testing is an integral part of the management of patients infected with HIV. The traditional Sanger sequencing method is capable of detecting drug resistant mutations (DRMs) that make up at least 10-15% of the viral quasispecies population. Newer next generation sequencing technologies have a greater sensitivity for the detection of minority variant DRMs down to around 1% of the population. OBJECTIVES: Here NGS sequencing on the Vela Diagnostics automated next generation sequencing platform was evaluated and compared to the currently used Sanger sequencing method. STUDY DESIGN: Sequences from both methods were obtained from a total of 79 patients, with a range of subtypes (CRF01_AE, A1/G, A1/CRF01_AE, A1/CRF02_AG, A1, A, B, C, CRF01_AG, CRF 06_CPX, D, G, B/G, CRF 57_BC/C, G/CRF 02_AG and CRF 14_BG/G) and viral loads (2.43-7 log10 copies/ml). RESULTS: A high concordance was seen between the two methods for subtyping (96%) and majority variant detection (97.9%). NGS sequencing detected more variants and DRMs than Sanger sequencing. Of the 76 patient samples 86% (n = 66) had identical drug resistance reports. From the ten discrepant reports, nine had extra DRMs detected by NGS sequencing and all discrepancies were seen for NRTI and NNRTI antiviral resistance. CONCLUSIONS: This study demonstrated a good performance of the NGS method for HIV-1 genotyping compared to the Sanger sequencing method for detection of majority variants, however the reproducibility for the detection of minority variants was sub-optimal. Adoption of an NGS sequencing approach has the potential to improve the clinical management of HIV-infected patients.

Comparative evaluation of 2 automated molecular systems for the detection of HSV-1 and 2 from genital swab specimens.

BD ProbeTec HSV 1 & 2 Qx (on the BD Viper XTR) and the Aptima HSV 1 & 2 (on the Hologic Panther) assays were compared. Of 257 clinical samples, 96.6% positive and 88.5% negative results were concordant, respectively. The BD assay was more sensitive than the Aptima, but the latter was more specific.

Evaluating the aptima HIV-1 quant Dx, HCV quant Dx and HBV quant assays against the Abbott HIV-1, HCV and HBV RealTime assays.

BACKGROUND: Viral load measurement is routine for the management of patients infected with HIV, HCV or HBV, using sensitive, quantitative, nucleic acid amplification tests (NAATs). However, platforms are not equivalent in terms of turnaround time, random access capability and operator hands-on time. OBJECTIVES AND STUDY DESIGN: We compared the performance of the Hologic Panther transcription-mediated amplification based Aptima assays for HIV, HBV and HCV viral load measurement with the corresponding Abbott m2000 RealTime assays. All Aptima assays were run according to the manufacturer's instructions, on archived patient samples for HIV (n = 251 including subtypes 01_AE, A (A1), B, BG recombinant, C CRF02_AG and G), HBV (n = 117 including genotypes A, B, C and D) and HCV (n = 82 including genotypes 1, 2, 3 and 4). Additional testing was performed with WHO international standards and expanded HIV-1 subtype and HCV NIBSC genotype panels. RESULTS: The Hologic Panther Aptima assays for HIV-1 RNA, HBV DNA and HCV RNA viral load measurements performed equivalently to the Abbott m2000 RealTime assays. The performance of the Aptima assays was also acceptable on the WHO and NIBSC standards and subtype/genotype panels. CONCLUSIONS: The Aptima Panther platform offers equivalent clinical performance for the viral load measurement of these three viruses, with the added benefits of reduced turnaround time, random access capability and reduced 'hands-on time' for staff, resulting in a potentially superior workflow for diagnostic laboratories.

Hepatitis B Virus Immunization and Neonatal Acquisition of Persistent Infection in England and Wales.

Background: It is believed that between 2% and 5% of infants born to hepatitis B virus (HBV)-infected mothers at a high risk of perinatal transmission will become persistently infected despite immunization starting at birth. We investigated factors associated with breakthrough infections. Methods: Sixty-nine samples from HBV-infected infants born between 2003 and 2015 were tested for HBV serological and molecular markers. Sequencing and epitope phenotyping were used to investigate alterations in hepatitis B surface antigen (HBsAg) sequence and antigenicity in infants and in mothers known to have transmitted and not to have transmitted virus to their infants. Results: Vaccine/hepatitis B immune globulin uptake was complete in the majority of HBV-infected infants. A minority (8 [12%]) had detectable plasma antibody to HBsAg at 12 months. Twenty-five of 68 (37%) infants harbored a virus with amino acid changes in the HBsAg "a" determinant, of which 13 displayed altered HBsAg antigenicity. Viral load was 30-fold higher in maternal samples from those who transmitted. Conclusions: Our data provide evidence to suggest that immune selection drives change at mother-infant transmission, resulting in the alteration of HBsAg antigenicity. These changes may play a role in immunization failure, but other factors including viral load may be more important. Continued monitoring of vaccine efficacy is essential.

Molecular epidemiology of newly acquired hepatitis C infections in England 2008-2011: genotype, phylogeny and mutation analysis.

BACKGROUND: Analysis of laboratory testing data collected through the Sentinel Surveillance programme has provided a method for identifying individuals who have recently acquired their hepatitis C virus (HCV) infection. Access to samples from these individuals provided a rare opportunity to undertake molecular characterization studies. OBJECTIVES: To describe the epidemiology and genetic diversity of hepatitis C in recent seroconverter infections and to predict how this will impact on HCV treatment and control. STUDY DESIGN: One hundred and forty seven samples were available from individuals, identified to have recently acquired their HCV infection. Genotype determination with additional phylogenetic analysis was carried out on NS5B sequences. Analysis across the NS3 region investigated the presence of antiviral resistance mutations. Where possible, molecular data was linked to demographic and risk/behavioural factor information. RESULTS: The majority of new infections occurred in males with a mean age of 37 years. The most commonly observed genotypes were 1a (49%) and 3a (42%) and injecting drug use (58%) was the most common risk factor. Genotype distribution differed between persons who inject drugs and those with other risk factors suggesting two possible epidemics. Phylogenetic analysis indicated possible transmission networks within specific risk groups. Amino acid changes associated with antiviral resistance were noted in the NS3 region in some samples. CONCLUSIONS: Continued surveillance of linked molecular, virological, demographic and epidemiological information on recently acquired infections will contribute to understanding the on-going HCV epidemic in England.