The effect of mouse strain on herpes simplex virus type 1 (HSV-1) infection of the central nervous system (CNS).

BACKGROUND: Mice infected with HSV-1 can develop lethal encephalitis or virus induced CNS demyelination. Multiple factors affect outcome including route of infection, virus and mouse strain. When infected with a sub-lethal dose of HSV-1 strain 2 via the oral mucosa, susceptible SJL/J, A/J, and PL/J mice develop demyelinating lesions throughout the brain. In contrast, lesions are restricted to the brainstem (BST) in moderately resistant BALB/c mice and are absent in resistant BL/6 mice. The reasons for the strain differences are unknown. METHODS: In this study, we combine histology, immunohistochemistry, and in-situ hybridization to investigate the relationship between virus and the development of lesions during the early stage (< 24 days PI) of demyelination in different strains of mice. RESULTS: Initially, viral DNA and antigen positive cells appear sequentially in non-contiguous areas throughout the brains of BALB/c, SJL/J, A/J, and PL/J mice but are restricted to an area of the BST of BL/6 mice. In SJL/J, A/J, and PL/J mice, this is followed by the development of 'focal' areas of virus infected neuronal and non-neuronal cells throughout the brain. The 'focal' areas follow a hierarchical order and co-localize with developing demyelinating lesions. When antigen is cleared, viral DNA positive cells can remain in areas of demyelination; consistent with a latent infection. In contrast, 'focal' areas are restricted to the BST of BALB/c mice and do not occur in BL/6 mice. CONCLUSIONS: The results of this study indicate that susceptible mouse strains, infected with HSV-1 via the oral mucosa, develop CNS demyelination during the first 24 days PI in several stages. These include: the initial spread of virus and infection of cells in non-contiguous areas throughout the brain, the development of 'focal' areas of virus infected neuronal and non-neuronal cells, the co-localization of 'focal' areas with developing demyelinating lesions, and latent infection in a number of the lesions. In contrast, the limited demyelination that develops in BALB/c and the lack of demyelination in BL/6 mice correlates with the limited or lack of 'focal' areas of virus infected neuronal and non-neuronal cells in these two strains.

The relationship between respiratory viral loads and diagnosis in children presenting to a pediatric hospital emergency department.

BACKGROUND: respiratory viral infections account for a considerable proportion of pediatric emergency room visits. Illnesses range in severity from mild upper respiratory tract infections to serious lower respiratory tract infections (LRTI). The relationship between viral load and specific viruses to clinical diagnosis made by physicians in this setting is poorly understood. METHODS: we applied a real-time, quantitative polymerase chain reaction (qPCR) panel for 13 common respiratory viruses to 195 frozen, archival nasopharyngeal aspirate specimens obtained from symptomatic children ≤ 4 months of age presenting to the emergency room. Mean total viral load and number of viruses per archival nasopharyngeal aspirate specimen were compared between LRTI (n = 70) and non-LRTI (1 or more of upper respiratory tract infection, fever, or cough) (n = 125), as were yield and concordance of qPCR results to viral culture/direct fluorescence assay (DFA). RESULTS: children with LRTI had significantly increased total viral load and harbored more viruses than the non-LRTI group. Respiratory syncytial virus-A and -B were significantly associated with LRTI, and parainfluenza virus-1 with non-LRTI. Individual loads of parainfluenza virus-2 and human rhinovirus were increased in LRTI versus non-LRTI. Quantitative PCR yielded more viruses (including coinfections, where a "dominant virus" was typically identified) than viral culture/DFA and documented nucleic acid from pathogens not tested by culture/DFA including human rhinovirus; coronaviruses -OC43, -229E, and -NL63; and metapneumovirus. CONCLUSIONS: in symptomatic children presenting to the emergency room, total viral load is related to clinical diagnosis; specific viruses are associated with particular clinical diagnoses, and qPCR has a higher yield than other viral diagnostic methods.

Is the healthy middle ear a normally sterile site?

OBJECTIVE: To systematically evaluate the presumption that the healthy middle ear becomes colonized with organisms via the patent eustachian tube using modern microbiologic techniques. STUDY DESIGN: Sterile saline washings were obtained from the middle ear of patients in a prospective fashion. SETTING: Tertiary/quaternary referral centers. PATIENTS: Pediatric and adult patients undergoing cochlear implantation surgery. INTERVENTION(S): Standard bacterial and viral cultures, and nucleic acid amplification techniques. MAIN OUTCOME MEASURE(S): Identification of organisms. RESULTS: Specimens were obtained from 13 children and 9 adults. No organisms were identified in any of the specimens, either through standard culture or PCR testing. CONCLUSION: The presumption that the healthy middle ear is colonized by bacteria from the nasopharynx is unsubstantiated.

Viridans streptococci bacteremia in children with malignancy: relevance of species identification and penicillin susceptibility.

Our aim was to determine the significance of species identification and penicillin susceptibility of viridans streptococci in children with malignancies. Streptococcus mitis accounted for 58% of invasive viridans streptococcal infections of which 51% were penicillin-nonsusceptible. There was no significant association between species or penicillin susceptibility pattern and clinical presentation or outcome.

Detection and genotyping of varicella-zoster virus by TaqMan allelic discrimination real-time PCR.

A proportion of individuals vaccinated with live attenuated Oka varicella-zoster virus (VZV) vaccine subsequently develop attenuated chicken pox and/or herpes zoster. To determine whether postvaccination varicella infections are caused by vaccine or wild-type virus, a simple method for distinguishing the vaccine strain from wild-type virus is required. We have developed a TaqMan real-time PCR assay to detect and differentiate wild-type virus from Oka vaccine strains of VZV. The assay utilized two fluorogenic, minor groove binding probes targeted to a single nucleotide polymorphism in open reading frame 62 that distinguishes the Oka vaccine from wild-type strains. VZV DNA could be genotyped and quantified within minutes of thermocycling completion due to real-time monitoring of PCR product formation and allelic discrimination analysis. The allelic discrimination assay was performed in parallel with two standard PCR-restriction fragment length polymorphism (RFLP) methods on 136 clinical and laboratory VZV strains from Canada, Australia, and Japan. The TaqMan assay exhibited a genotyping accuracy of 100% and, when compared to both PCR-RFLP methods, was 100 times more sensitive. In addition, the method was technically simpler and more rapid. The TaqMan assay also allows for high-throughput genotyping, making it ideal for epidemiologic study of the live attenuated varicella vaccine.