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Human cytomegalovirus strains associated with congenital and perinatal infections
D E Trincado1, G M Scott, P A White
1Virology Division, Department of Microbiology, South Eastern Area Laboratory Services, The Prince of Wales Hospital, Randwick, Australia.
Abstract:
The genotypes of human cytomegalovirus (HCMV) isolates from pediatric patients differs from those of infected adults in Australia. Genotypes were determined by PCR amplification of glycoprotein B (gB) sequences, with subsequent analysis by restriction fragment length polymorphism, single-stranded conformation polymorphism, heteroduplex mobility analysis and direct DNA sequencing. Restriction fragment length polymorphism analysis of gB showed genotypes gB1 (39%) and gB3 (30%) were more prevalent in infected children and two new genotypes (gB6 and gB7) were found. Single-stranded conformation polymorphism was used to group isolates into 22 further subtypes and suggested longitudinal co-infection or viral mutation was occurring over time. Heteroduplex mobility analysis was found to be the most accurate and concise of the four methods used for genotyping HCMV isolates. DNA sequencing was used to confirm the results obtained from heteroduplex mobility analysis, and identified two isolates that were incorrectly genotyped by restriction fragment length polymorphism analysis. Heteroduplex mobility analysis efficiently genotyped all samples and allowed estimation of sequence variation between isolates. These data suggest certain gB genotypes are associated more commonly with childhood infections, and these differ from strains associated with invasive disease in HIV patients.
Insights
Human cytomegalovirus (HCMV) genotypes in Australian children differ from adults, with specific gB types more common in pediatric infections. Heteroduplex mobility analysis proved most effective for accurate HCMV genotyping.
Area of Science:
- Virology
- Genetics
- Infectious Diseases
Background:
- Human cytomegalovirus (HCMV) is a common pathogen with diverse genotypes.
- Understanding HCMV genotype distribution is crucial for disease management, particularly in pediatric populations.
- Previous studies have not fully characterized HCMV gB genotypes in Australian pediatric patients.
Purpose of the Study:
- To determine the genotypes of human cytomegalovirus (HCMV) isolates from pediatric patients in Australia.
- To compare these genotypes with those found in infected adults.
- To evaluate the accuracy and efficiency of different genotyping methods for HCMV.
Main Methods:
- Polymerase chain reaction (PCR) amplification of glycoprotein B (gB) sequences from HCMV isolates.
- Analysis using restriction fragment length polymorphism (RFLP), single-stranded conformation polymorphism (SSCP), heteroduplex mobility analysis (HMA), and direct DNA sequencing.
- Comparison of the four genotyping techniques for accuracy, conciseness, and efficiency.
Main Results:
- HCMV gB genotypes in pediatric patients differed from those in adults.
- Genotypes gB1 (39%) and gB3 (30%) were prevalent in children; novel genotypes gB6 and gB7 were identified.
- Heteroduplex mobility analysis (HMA) was the most accurate and concise method, confirming results and identifying discrepancies with RFLP.
Conclusions:
- Specific HCMV gB genotypes are more commonly associated with childhood infections in Australia.
- These pediatric-associated genotypes differ from those linked to invasive disease in HIV patients.
- Heteroduplex mobility analysis is a highly effective method for HCMV genotyping and assessing sequence variation.