To assess the concordance of high-risk HPV (HR-HPV) testing with the Alinity assay on cervical samples collected with diverse collection/storage protocols (ThinPrep, SurePath, Cervicollect) and to assess inter-assay concordance of HR-HPV testing of cervical cell specimens with Alinity m HR HPV assay (Alinity) vs cobas® 4800 HPV assay (cobas).Specimens were obtained from 560 women attending a Women's Health clinic. Two specimens were obtained from each woman with combinations of two of the three collection devices and aliquots were tested by the two assays.Alinity showed an agreement of 93.9%, Kappa = 0.89 (263/280) between ThinPrep and SurePath specimens; 97.5%, Kappa = 0.95 (347/356) and 92.9%, Kappa = 0.85 (104/112) between ThinPrep and SurePath aliquots taken before or after cytology processing, respectively. Cervi-Collect specimens showed an agreement of 94.6%, Kappa = 0.89 (265/280) with ThinPrep specimens. Compared to cobas, Alinity showed agreements of 94.3%, Kappa = 0.88 (395/419) and 91.8%, Kappa = 0.82 (257/280) between ThinPrep and SurePath specimens, respectively. Alinity and cobas detected genotypes 16/18 and other high-risk HPV types at similar rates and showed similar correlations with cytology grades.Compared to cobas, Alinity performed equally well for detecting HPV in cervical specimens obtained with ThinPrep and SurePath. The Cervi-Collect device compared well to the other collection methods. Alinity is a reliable assay for simultaneous detection of HPV-16/18 and other high-risk genotypes in cervical specimens.
Abstract The outer membrane (OM) of Gram-negative bacteria is a unique asymmetric lipid bilayer in which the outer leaflet is composed of lipopolysaccharide (LPS) and the inner leaflet is formed by glycerophospholipid (GPL). The OM plays a fundamental role in protecting Gram-negative bacteria from harsh environments and toxic compounds. The transport and assembly pathways for phospholipids of bacterial OM are unknown. Cardiolipin (CL) plays an important role in OM biogenesis and pathogenesis, and the inner membrane (IM) protein PbgA, containing five transmembrane domains and a globular domain in periplasm has been recently identified as a CL transporter from the IM to the OM with an unknown mechanism. Here we present the first two crystal structures of soluble periplasmic globular domain of PbgA from S. typhimurium and E. coli , which revealed that the globular domains of PbgA resemble the structures of the arylsulfatase protein family and contains a novel core hydrophobic pocket that may be responsible for binding and transporting CLs. Our structural and functional studies shed an important light on the mechanism of CL transport in Gram-negative bacteria from the IM to the OM, which offers great potential for the development of novel antibiotics against multi-drug resistant bacterial infections.
HIV is characterized by a high degree of genetic diversity, presenting a challenge for the development of assays for initial diagnosis and subsequent monitoring of therapy response. Alinity m HIV-1 was developed on the Alinity m System, a fully automated, random/continuous access analyzer, to achieve accurate quantitation across groups, subtypes and circulating recombinant forms (CRF), concurrent HIV-1 confirmation and viral load monitoring in plasma, confirmation in serum.
Methods
Abbott's Global Surveillance program was utilized to identify the most conserved target regions across HIV-1 variants. The assay targets two HIV-1 genomic regions and utilizes partially double-stranded probes, RNA-specific sample preparation chemistry, unit-dose lyophilized amplification reagents, and patented ReadiFlex™ sample processing logistics to deliver a time-to-first-result of 115 minutes. The Alinity m HIV-1 assay was evaluated for key performance attibutes.
Results
Alinity m HIV-1 demonstrated linearity from 10 to 20,000,000 Copies/mL and demonstrated a within-laboratory SD of ≤0.13 Log Copies/mL from 2.3 to 7.4 Log Copies/mL. Probit analysis demonstrated that the assay detected HIV-1 RNA with 95% probability at 13.88 Copies/mL using 3rd WHO HIV-1 Standard (subtype B). The assay exhibited ≥95% detection for HIV-1 group M subtypes, groups O and N at 20 Copies/mL. Correlation between Alinity m HIV-1 and Abbott RealTime HIV-1 assays demonstrated a mean bias of -0.03 Log Copies/mL (95% CI: -0.05 to 0.00). Confirmatory method agreement between Alinity m HIV-1 assay and comparator HIV-1 assay was 100%.
Conclusion
The Alinity m HIV-1 assay utilizes a state of the art instrument system and dual-target assay design to deliver highly sensitive detection of diverse HIV-1 groups/subtypes and accurate quantitation across a wide dynamic range while facilitating rapid turnaround time (115 minutes) and workflow flexibility. By providing confirmation and baseline viral load measurement in one test, the assay reduces the number of steps required for initial diagnosis of infection.
Abstract Background Semen cryopreservation is a critical tool for breed improvement and preservation of biodiversity. However, instability of sperm freezability affects its application. The Mediterranean buffalo is one of the river-type buffaloes with the capacity for high milk production. Until now, there is no specific cryopreservation system for Mediterranean buffalo, which influences the promotion of excellent cultivars. To improve the semen freezing extender used in cryopreservation of Mediterranean buffalo, different protein datasets relating to freezability sperm were analyzed by iTRAQ-based proteomics. This study will be beneficial for further understanding the sperm freezability mechanism and developing new cryopreservation strategy for buffalo semen. Results 2652 quantified proteins were identified, including 248 significantly differentially expressed proteins (DEP). Gene Ontology (GO) analysis indicated that many these were mitochondrial proteins, enriched in the molecular function of phospholipase A2 activity and enzyme binding, and biological processes of regulation of protein kinase A signaling and motile cilium assembly. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis identified 17 significant pathways, including oxidative phosphorylation (OXPHOS). Furthermore, 7 DEPs were verified using parallel reaction monitoring or western blot, which confirmed the accuracy of the iTRAQ data. Peroxiredoxin 6 (PRDX6), which was expressed 1.72-fold higher in GFE compared to PFE sperm, was selected to explore the function in sperm freezability by adding recombinant PRDX6 protein into the semen freezing extender. The results showed that the motility, mitochondrial function and in vitro fertilization capacity of frozen-thawed sperm were significantly increased, while the oxidation level was significantly decreased when 0.1mg/L PRDX6 was added compared with blank control. Conclusions Above results revealed the metabolic pattern of freezability of Mediterranean buffalo sperms was negatively associated with OXPHOS, and PRDX6 had protective effect on cryo-damage of frozen-thawed sperms.
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