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Seven Australian infectious bronchitis viruses incorporated radioactive amino acid label during growth in two types of cell culture. From purified virions, four major polypeptides of molecular weights 91,000 (GP91), 51,000 (VP51), 25,000 (GP25) and 23,000 (VP23) were separated by PAGE. Two minor polypeptides of molecular weights 210,000 (GP210) and 185,000 (GP185) were also identified. 3H-glucosamine was incorporated during growth of virus into GP210, GP185, GP91 and GP25. Bromelain digestion of virions removed GP210, GP185 and GP91, leaving VP51, GP25 and VP23 in a large sub viral particle. All seven viruses, which were of six serological subtypes, formed apparently identical polypeptide profiles.

Viral isolation is the standard method for the detection of herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) in clinical specimens. This study describes the development of a type-specific polymerase chain reaction (PCR) assay for detection and typing of HSV-1 and HSV-2, and a comparison of its sensitivity with that of isolation in a clinical setting. Specimens from patients presenting with genital ulcers were tested for the presence of HSV by both methods. Oligonucleotide primers were selected to enable type-specific amplification of HSV-1 and HSV-2 DNA. Conditions were optimized to allow detection and typing from a single reaction tube using a multi-primer PCR method. When compared with PCR, the sensitivity of isolation was 67% and the specificity 97%. This protocol allowed rapid, sensitive and accurate detection and typing of HSV with a single PCR assay.

Viral isolation is the standard method for the detection of herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) in clinical specimens. This study describes the development of a type-specific polymerase chain reaction (PCR) assay for detection and typing of HSV-1 and HSV-2, and a comparison of its sensitivity with that of isolation in a clinical setting. Specimens from patients presenting with genital ulcers were tested for the presence of HSV by both methods. Oligonucleotide primers were selected to enable type-specific amplification of HSV-1 and HSV-2 DNA. Conditions were optimized to allow detection and typing from a single reaction tube using a multi-primer PCR method. When compared with PCR, the sensitivity of isolation was 67% and the specificity 97%. This protocol allowed rapid, sensitive and accurate detection and typing of HSV with a single PCR assay.

No abstract