The relationship between human papillomavirus (HPV) DNA in the genital mucosa and serum IgG to HPV-16, -18, and -6 was studied in a cohort of 588 college women. Among women with incident HPV infections, 59.5%, 54.1%, and 68.8% seroconverted for HPV-16, -18, or -6, respectively, within 18 months of detecting the corresponding HPV DNA. Transient HPV DNA was associated with a failure to seroconvert following incident HPV infection; however, some women with persistent HPV DNA never seroconverted. Antibody responses to each type were heterogeneous, but several type-specific differences were found: seroconversion for HPV-16 occurred most frequently between 6 and 12 months of DNA detection, but seroconversion for HPV-6 coincided with DNA detection. Additionally, antibody responses to HPV-16 and -18 were significantly more likely to persist during follow-up than were antibodies to HPV-6.

Testing for HPV has higher sensitivity but lower specificity than thin-layer Pap screening. In some settings, particularly where screening intervals are long or haphazard, screening for HPV DNA may be a reasonable alternative to cytology-based screening of reproductive-age women.

Conventional fluorescent-antibody (FA) methods were compared to real-time PCR assays for detection of respiratory syncytial virus (RSV), influenza virus type A (FluA), parainfluenza virus types 1, 2, and 3 (PIV1, PIV2, and PIV3), human metapneumovirus (MPV), and adenovirus (AdV) in 1,138 specimens from children with respiratory illnesses collected over a 1-year period. At least one virus was detected in 436 (38.3%) specimens by FA and in 608 (53.4%) specimens by PCR (P < 0.001). Specimen quality was inadequate for FA in 52 (4.6%) specimens; 13 of these (25%) were positive by PCR. In contrast, 18 (1.6%) specimens could not be analyzed by PCR; 1 of these was positive by FA. The number of specimens positive only by PCR among specimens positive by PCR and/or FA was 18 (7.0%) of 257 for RSV, 18 (13.4%) of 134 for FluA, 25 (64.1%) of 39 for PIV1, 8 (88.9%) of 9 for PIV2, 17 (30.1%) of 55 for PIV3, and 101 (76.5%) of 132 for AdV. MPV was detected in 6.6% of all specimens and in 9.5% of the 702 specimens negative by FA. The mean number of virus copies per milliliter in specimens positive by both PCR and FA was significantly higher, at 6.7 ؋ 10 7 , than that in specimens positive only by PCR, at 4.1 ؋ 10 4 (P < 0.001). The PCR assays were significantly more sensitive than FA assays for detecting respiratory viruses, especially parainfluenza virus and adenovirus. Use of real-time PCR to identify viral respiratory pathogens in children will lead to improved diagnosis of respiratory illness.Accurate detection of respiratory viruses is important to guide antiviral therapy, prevent nosocomial spread, provide surveillance, and in some cases, decrease hospital costs and lengths of stay (1, 2, 11, 21). By using standard laboratory methods, such as staining with fluorescent antibodies (FA) and isolation by culture, viruses have been detected in 13 to 45% of children with symptoms of respiratory illness (3,8,12,22,28). Disadvantages of FA include requiring multiple reagents which may vary in sensitivity, potential variability in technical reading, and the need for an adequate number of cells to examine each specimen. Several studies have shown that PCR methods appear to be more sensitive than FA and culture for the diagnosis of acute respiratory virus infections (8,22,23,24,26,28). PCR is less affected by specimen quality and transport and provides an objective interpretation of results. Real-time PCR technology, which combines nucleic acid amplification with amplicon detection, provides results more quickly than conventional PCR, has in some cases shown improved sensitivity compared to conventional PCR, and provides a uniform platform for quantifying both single and multiple pathogens in a single sample (4,7,18).In this study, separate quantitative real-time reverse transcription (RT)-PCR assays were used to detect six RNA viruses, including respiratory syncytial virus (RSV), influenza virus type A (FluA), parainfluenza virus types 1, 2, and 3 (PIV1, PIV2, and PIV3), and human metapneumovirus (MPV). A quantitative real-time PCR a...

Background Molecular testing for viral pathogens has resulted in increasing detection of multiple viruses in respiratory secretions of ill children. The clinical impact of multiple-virus infections on clinical presentation and outcome is unclear. Objectives To compare clinical characteristics and viral load between children with multiple-virus versus single-virus illnesses. Patients/Methods 893 residual nasal wash samples from children treated for respiratory illness at Children's Hospital, Seattle, from September 2003 through September 2004 were evaluated by quantitative PCR for respiratory syncytial virus (RSV), human metapneumovirus (hMPV), influenza (Flu), parainfluenza (PIV), adenoviruses (AdV), and coronaviruses (CoV). Illness severity and patient characteristics were abstracted from medical charts. Results Coinfections were identified in 103 (18%) of 566 virus-positive samples. Adenovirus was most commonly detected in coinfections (52%), followed by coronavirus (50%). Illnesses with a single virus had increased risk of oxygen requirement (p=0.02), extended hospital stays (p=0.002), and admissions to the inpatient (p=0.02) or intensive care units (p=0.04). For Adv and PIV-1, multiple-virus illnesses had a significantly lower viral load (log10 copies/mL) than single-virus illnesses (4.2 vs 5.6, p=0.007 and 4.2 vs 6.9, p<0.001, respectively). RSV, Flu-A, PIV-3, and hMPV viral loads were consistently high whether or not another virus was detected. Conclusions Illnesses with multiple-virus detections were correlated with less severe disease. The relationship between viral load and multiple-virus infections was virus specific, and this may serve as a way to differentiate viruses in multiple-virus infections.

Background Little is known about HBoV persistence and shedding and the association between HBoV detection and the onset and resolution of respiratory symptoms. Methods We performed HBoV testing on nasal swabs from a prospective, longitudinal study of respiratory illness in 119 children attending daycare. Results HBoV was detected in 70 children (59%), and in 106 (33%) of the 318 study illnesses. Another virus was detected in 76 HBoV+ illnesses (72%). Extended and intermittent shedding was observed, with consistent HBoV detection documented for up to 75 days. HBoV was detected in 20 of 45 asymptomatic enrollment samples (44%) and HBoV prevalence and viral load did not differ significantly between children with and without symptoms at enrollment. HBoV+ illnesses were longer than HBoV- illnesses (OR: 2.44 for symptoms > 7 days, 95% C.I. 1.41, 4.22) and illnesses with HBoV viral load ≥ 4 log-copies/mL required a visit to a health care provider more often than HBoV-illnesses (OR: 1.64, 95% C.I.: 1.02, 2.64). Conclusion HBoV was more common in illnesses with greater severity. However, detection of HBoV was not associated with the presence of respiratory illness or with specific respiratory symptoms in this prospective study of infants and toddlers attending center-based daycare.