Restriction Endonuclease Fingerprinting of Herpes Simplex Virus DNA: A Novel Epidemiological Tool Applied to a Nosocomial Outbreak

Abstract

In a blind study, 14 isolates of herpes simplex virus type 1 (HSV-1) that included nine isolates from a temporal cluster of HSV infections in a hospital Pediatric Intensive Care Unit and five unrelated isolates were analyzed by digestion of their DNA with four restriction endonucleases. These enzymes (HsuI, BglII, EcoRI, and HpaI) cleave the DNA in about 52 sites. To date, at least 16 sites have been found to be variable in the sense that they may be present or absent independently of any other cleavage site. This characteristic is stable, and no change was observed on serial propagation of the strains in culture or following repeated isolation, as long as 12 years apart, from humans. Analyses of the isolates readily discriminated between those belonging to the temporal cluster of hospital infections and the unrelated strains. They also showed that there were two independent introductions of HSV-1 into the Pediatric Intensive Care Unit resulting in two clusters of epidemiologically related infections. This type of analysis has the potential of becoming a powerful tool for tracing the spread of HSV-1 and very likely of other herpesviruses in the human population.

Retrieval of Latent Herpes Simplex Virus Type 1 Genetic Information from Murine Trigeminal Ganglia by Superinfection with Heterotypic Virus in vivo

“…1 x 108 cells) were infected with 0.1 p.f.u, of HSV per cell in 10 ml of medium at 37 °C for 1 h. Eagle's MEM (20 ml) was then added to each bottle, which was incubated at 37 °C for 48 h. The infected cells were scraped off with a rubber policeman, washed twice in phosphate-buffered saline (PBS), and then collected by low-speed centrifugation (800 g for 10 min at 4 °C). HSV DNA was extracted by a modification of methods described previously (Green et al, 1971 ;Buchman et aL, 1978 ;Lonsdale et al, 1979 ;Pignani et al, 1979). Five ml of lysing solution (0.25 ~ Triton X-100, 10 mM-EDTA, 10 mM-Tris HC1 pH 7.9) was added to the infected cells.…”

“…The pellet was washed twice with 70% ethanol, dried under vacuum, and dissolved in 200 Ixl Tris-EDTA buffer. Viral DNA was then digested as described by Buchman et al (1978). Restriction endonucleases BamHI, HindIII and EcoRI were purchased from Boehringer, and for cleavage the appropriate buffer recommended by the supplier was used for each enzyme.…”

Restriction Endonuclease DNA Analysis of Leptospira interrogans Serovars Icterohaemorrhagiae and Copenhageni

“…Originally, restriction endonuclease DNA analysis was developed for viral DNA analysis (3,4). The application of restriction endonuclease DNA analysis to the classification of leptospires was first proposed by Marshall et al (11) in 1981 .…”

Antibody Response, Recurrence Patterns and Subsequent Herpes Simplex Virus Type 2 (HSV-2) Re-infection Following Initial HSV-2 Infection of Guinea-pigs: Effects of Acyclovir

“…Confluent monolayers of Vero cells, washed twice with phosphate-free Eagle's ME M supplemented with 2 ~ foetal calf serum, were inoculated with viral isolates at a multiplicity of infection of 1 p.f.u./cell. HSV DNA was labelled with 32p i (50 ~tCi/ml), extracted and prepared for restriction analysis by a modification of methods previously reported (Buchman et al, 1978;Lonsdate, 1979). Restriction endonuclease analysis was performed with BamHI under the conditions specified by the manufacturer after preliminary experiments had determined that this enzyme readily differentiated the strains MS and 333.…”