Effect of glacial acetic acid treatment of liquid-based cytology collections on performance of Cervista HPV HR for detection of high-risk human papillomavirus.

Inadequate cervical cytological analysis can be facilitated by glacial acetic acid (GAA) treatment of primary liquid-based collections to remove mucus, erythrocytes, inflammatory cells, and cellular debris. In the context of a commercial human papillomavirus (HPV) hybridization assay performed on 465 tandem specimens with and without GAA treatment, we show that GAA treatment significantly reduces genomic DNA content (P < 0.0001) and creates an increased potential for indeterminate and false-negative results. In the context of cytological workflow, laboratories should consider providing a specimen aliquot for HPV DNA detection prior to GAA treatment.

Hamster and murine models of severe destructive Lyme arthritis.

Arthritis is a frequent complication of infection in humans with Borrelia burgdorferi. Weeks to months following the onset of Lyme borreliosis, a histopathological reaction characteristic of synovitis including bone, joint, muscle, or tendon pain may occur. A subpopulation of patients may progress to a chronic, debilitating arthritis months to years after infection which has been classified as severe destructive Lyme arthritis. This arthritis involves focal bone erosion and destruction of articular cartilage. Hamsters and mice are animal models that have been utilized to study articular manifestations of Lyme borreliosis. Infection of immunocompetent LSH hamsters or C3H mice results in a transient synovitis. However, severe destructive Lyme arthritis can be induced by infecting irradiated hamsters or mice and immunocompetent Borrelia-vaccinated hamsters, mice, and interferon-gamma- (IFN-γ-) deficient mice with viable B. burgdorferi. The hamster model of severe destructive Lyme arthritis facilitates easy assessment of Lyme borreliosis vaccine preparations for deleterious effects while murine models of severe destructive Lyme arthritis allow for investigation of mechanisms of immunopathology.

Clinical laboratory evaluation of Invader® chemistry and hybrid capture for detection of high-risk human papillomavirus in liquid-based cytology specimens.

A laboratory-developed test for high-risk human papillomavirus (HPV) that utilizes the Invader technology (Hologic, Madison, WI, USA) was compared to hybrid capture (Digene, Gaithersburg, MD, USA). A total of 342 ThinPrep specimens were de-identified following cytology screening (10 high-grade squamous intraepithelial lesions, 14 low-grade squamous intraepithelial lesions, 199 atypical squamous cells of undetermined significance, 119 normal). DNA was manually extracted prior to interrogation with Invader. Hybrid capture/Invader discrepancies were subject to HPV sequencing. One sample (0.3%) was indeterminate by Invader due to low genomic DNA content. Concordance of 341 available tandem hybrid capture/Invader results occurred at a rate of 91.5%. Differences in HPV detection rate between the 2 assays were not statistically significant (P = 0.17). A propensity for false-positive hybrid capture result was confirmed by HPV sequencing in 83% of instances. Manual DNA extraction efficacy did not statistically differ between cytologic classifications (P ≥ 0.19). Invader detection of high-risk HPV is comparably sensitive and more specific than hybrid capture, providing an alternative for molecular HPV detection.

Gamma interferon inhibits production of Anti-OspA borreliacidal antibody in vitro.

The ability of a Lyme borreliosis vaccine to induce and maintain sustained levels of borreliacidal antibody is necessary for prolonged protection against infection with Borrelia burgdorferi. Vaccination against infection with B. burgdorferi could be improved by determining the mechanism(s) that influences the production of protective borreliacidal antibody. Borreliacidal antibody was inhibited in cultures of lymph node cells obtained from C3H/HeJ mice vaccinated with formalin-inactivated B. burgdorferi and cultured with macrophages and B. burgdorferi and treated with recombinant gamma interferon (rIFN-gamma). The suppression of production of outer surface protein A (OspA) borreliacidal antibody by rIFN-gamma was not affected by the time of treatment. In addition, treatment with rIFN-gamma inhibited the production of other anti-B. burgdorferi antibodies. By contrast, treatment of cultures of immune lymph node cells with anti-IFN-gamma marginally increased the production of borreliacidal antibody and enhanced the production of other antibodies directed against B. burgdorferi. These results show that IFN-gamma does not play a major role in the production of anti-OspA borreliacidal antibody. Additional studies are needed to determine which cytokine(s) will enhance production of borreliacidal antibody.