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Original Article |

Comparative Analysis of Detection Methods for Congenital Cytomegalovirus Infection in a Guinea Pig Model

Albert H. Park, MD; David Mann, MD; Marc E. Error, MD; Matthew Miller, BA; Matthew A. Firpo, PhD; Yong Wang, PhD; Stephen C. Alder, PhD; Mark R. Schleiss, MD
JAMA Otolaryngol Head Neck Surg. 2013;139(1):82-86. doi:10.1001/jamaoto.2013.1090.
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Objective  To assess the validity of the guinea pig as a model for congenital cytomegalovirus (CMV) infection by comparing the effectiveness of detecting the virus by real-time polymerase chain reaction (PCR) in blood, urine, and saliva.

Design  Case-control study.

Setting  Academic research.

Subjects  Eleven pregnant Hartley guinea pigs.

Main Outcome Measures  Blood, urine, and saliva samples were collected from guinea pig pups delivered from pregnant dams inoculated with guinea pig CMV. These samples were then evaluated for the presence of guinea pig CMV by real-time PCR assuming 100% transmission.

Results  Thirty-one pups delivered from 9 inoculated pregnant dams and 8 uninfected control pups underwent testing for guinea pig CMV and for auditory brainstem response hearing loss. Repeated-measures analysis of variance demonstrated no statistically significantly lower weight for the infected pups compared with the noninfected control pups. Six infected pups demonstrated auditory brainstem response hearing loss. The sensitivity and specificity of the real-time PCR assay on saliva samples were 74.2% and 100.0%, respectively. The sensitivity of the real-time PCR on blood and urine samples was significantly lower than that on saliva samples.

Conclusions  Real-time PCR assays of blood, urine, and saliva revealed that saliva samples show high sensitivity and specificity for detecting congenital CMV infection in guinea pigs. This finding is consistent with recent screening studies in human newborns. The guinea pig may be a good animal model in which to compare different diagnostic assays for congenital CMV infection.

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Figures

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Figure 1. Experimental process showing the animal outcomes for each step in the experimental design. ABR indicates auditory brainstem response; CMV, cytomegalovirus; ELISA, enzyme-linked immunosorbent assay; and PCR, polymerase chain reaction.

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Figure 2. Body mass of cytomegalovirus (CMV)–infected and CMV-uninfected pups. The mean body mass was determined at birth and at various postnatal time points for pups from sham-inoculated dams (CMV-negative dams) and from pups from CMV-inoculated dams (CMV-positive dams). Data represent the mean (SE).

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Figure 3. Auditory brainstem response thresholds in guinea pig pups. Auditory brainstem responses were determined at various decibels in a representative normal pup from a cytomegalovirus (CMV)–naive dam (CMV-negative pup) and in a representative pup with hearing loss from a CMV-inoculated dam (CMV-positive pup). In this comparison, the auditory brainstem response threshold in the CMV-negative pup was 20 dB, whereas the threshold in the CMV-positive pup was 55 dB. SPL indicates sound pressure level.

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Figure 4. Auditory brainstem response (ABR) thresholds in guinea pig pups by group. Although no overall ABR threshold difference is observed between cytomegalovirus (CMV)–positive and CMV-negative control groups (P > .05, 2-way analysis of variance), a subset of 6 CMV-positive animals showed threshold elevation of greater than 15 dB in at least 1 of the tested frequencies. Data represent the mean (SE). SPL indicates sound pressure level.

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