Hellenic Society of Virology

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Hellenic Virology

Volume 1, Number 1, 1996

Abstracts

Management of Herpes virus infectios in children.
M. Papagrigoriou-Theodoridou.
Pediatrics Department, University of Athens Medical School.
Hellenic Virology, 1996, 1(1): 13-16.
Herpes virus infections are of special interest for pediatrician because of the broad spectrum of cclinical manifestations, the possibilities of accurate diagnosis and effective treatment. For the prevention of the perinatal infections it is important that the pregnant woman at risk should be informed and have a proper follow up. The use of specific immunoglobulins has limited value while in the field of immunization the vaccine for the chickenpox has been proved to be safe and effective and is already in use in many countries. At present the vaccine is recommended for immunocompromised individuals. Acyclovir remains the drug of choice for herpes simplex and varicella-zoster virus infections. Acyclovir treatment should be reserved for defined subgroups. Oral acyclovir is not recommended routinely for varicella in healthy children and during late pregnancy. In AIDS patients herpesviruses may either act as opportunistic pathogens or the may interact to promote the pathogenicity of HIV. Recently herpesvirus infections are recognised as casual infecting organisms in new or old diseases.
Random peptide phage libraries and their use in Virology.
G. Koptopoulos.
Laboratory of Microbiology & Infectious Diseases, Veterinary Department, Aristotelian University of Thessaloniki.
Hellenic Virology, 1996, 1(1): 17-21.
The random peptide phage libraries are a new area of great scientific importance in biological sciences. They represent a vast mixture of filamentous phage clones and posses, on their surface, peptides which could operate as epitopes. By this method, which has been developed over the last 2-3 years, it is possible to produce certain epitopes, picked up by certain antibodies. It is an easy and very promissing procedure for in vitro antigen (epitope) preparation. They might be used as immunogens as well. Efforts have also been made lately to produce antibodies by using phage libraries.
Clinical enaluation of anti-p24 in HIV infection.
L. Kavallierou (1), N. Tsibourakis (1), J. Perdios (2), J. Stefanou (1), P. Gargalianos (2), P. Katsea (1), J. Kosmidis (2), N. Renieri (1).
(1) 3rd Peripheral Blood Center & (2) Special Infections Unit, Peripheral General Hospital of Athens.
Hellenic Virology, 1996, 1(1): 22-28.
Serum samples of 108 HIV seropositive subjects in different stages of HIV 1 infection were evaluated for p24 antigen (p24 Ag) and p24 antibodies (p24 Ab). ELISA assay was used for the determination of the results. During progressing disease, p24 Ag appeared with increasing concentrations while p24 Ab occured with decreasing titres. There was a strong correlation between PCP/S. Kaposi, elevated concentrations of p24 Ag and decreased titres of p24 Ab. AZT or ddI seems to permit the improvement of these immunological markers. Titres of anti-p24 seem to play an important role in clinical evaluation, therapy estimation and disease classification.
Virucidal effect of antiseptics and disinfectants on hepatitis A virus.
E. Biziagos.
Laboratory of Clinical Virology, Department of Medicine, University of Crete, Heraklion.
Hellenic Virology, 1996, 1(1): 33-37.
The effect of three antiseptics and three disinfectants on the infectivity of Hepatitis A virus (HAV) was examined. The CF53 viral strain, adapted to human hepatoma cell line PLC/PRF/5, in aqueous solution cotaining 100_g /ml of protein, was treated with an antiseptic or disinfectant at one appropiate concentration for 1,2,3,10,15 or 30 min. After the virucidal assays, compound and HAV were separated by gel filtration and viral infectivity titers were determined by cell culture titration. One antiseptic, Dakin's solution (5_g /ml free chlorine) and two disinfectants, 0.5% glutaraldehyde and sodium hypochlorite (4_g /ml free chlorine) proved effective against HAV. They reduced the virus infectivity titer by more than 5.4log10 after 1, 2 and 10 min respectively. 70% ethanol and chloramine T (4_g /ml free chlorine) reduced the HAV infectivity by 4log10 after a 15 min treatment and formaldehyde was ineffective at any treatment time used.
Molecular epidemiology of HPV infections of the anogenital track.
V. Labropoulou (1), E. Diakomanolis (2), K. Kalpaktsoglou (2), A. Balamotis (3), P. Mavromara (1)
(1) Molecular Virology Laboratory, Hellenic Pasteur Institute, (2) 1st Department of Obstretics and Gyneology, Alexandra Hospital, University of Athens, (3) Department of Dermatology, A. Sygros Hospital.
Hellenic Virology, 1996, 1(1): 38-41.
156 biopses from a broad spectrum of anogenital lesions were examined for presence of HPV DNA by Southern blot and PCR analysis. The results of this study are summarized as follows: In condylomata acuminata HPV DNA was detected in 95% of the biopses. The HPV types 6 and 11 and in particular the subtype HPV 6a were most frequently found. In low grade intraepithelial neoplasia lesions HPV DNA was detected in 72% of the bioopses and intermediate and high risk HPV types (6, 11, 16 18, 31, 33, 51, 53 and 66) were found. In high grade intrepithelial lesions HPV types 16 and 18 were detected in the majority of the biopses, whereas HPV 31, 33 and 51 were less frequently detected. Overall, 88% of the biopses has detecable HPV DNA. In biopses from invasive cancer, HPV DNA was detected in 93% of the cases. HPV 16 and HPV 18 were the predominant types. Uncharacterized HPV types were also detected in 22% of the biopses examined whereas HPV 18 appears to occur in high frequency. Two pontetially "novel" HPV types were also detected. Finally two HPV 16 variants were found and characterized in the material examined in this study.
Pathogenesis of three virus types on the olive fruit fly (Bactrocera (Dacus) oleae).
M. Anagnou-Veroniki.
Laboratory of Insect Microbiology & Pathology, Benakis Institute of Plant Pathology and National Institute of Agricultural Research
Hellenic Virology, 1996, 1(1): 42-45.
Pathogenesis caused by the artificial infection with viruses, of Reovirus, Iridovirus, and Picornavirus types, on the insect olive fruit fly (Bactrocera (Dacus) was studied aiming to the estimation of the possibility of using viruses in biological control of insects. The morphology of the viruses which settle in the tissues was studied and since virus morphogenesis, the kind of the cells and the cells and place where reproduce themselves was localized, the characterization of the disease was attempted.
Methodology development and detection of antiviral action of Hypercium L. on the Human Cytomegalovirus(HCMV).
S. Axarlis (1), M. Malamas (2), K. Demetzos (3), S. Mitakou (3), L. Skaltsounis (3), A. Mentis (1), N. Spyrou
(1) Hellenic Pasteur Institute, 127 Vas. Sofias Av. 157 21 Athens, (2) Department of Pharmacology, Medical School, University of Ionnina 451 10 Ioannina and (3) Department of Pharmacy University of Athens, Campus Zografou, 157 71 Athens.
Hellenic Virology, 1996, 1(1): 46-50.
In this preliminary work we developed a methodogy for screening different plants extracts for their antiviral activity against the Human Cytomegalovirus (HCMV). The same methodogy could by applied, with minor changes, in a large spectrum of other virus. The plant extracts first tested were from Hypericum perforatum L. We concluded that fraction IV of the Hypericum perforatum L. extracts that that had the strongest antiviral activity.
Development of methods for the detection of Hepatitis A from sludge and shellfishes.
H. Anastassiadou (1) and E. Biziagos (2).
(1) Institute of Nutritional Health, National Institute of Agricultural Research, Athens and (2) Clinical Virology Laboratory, Department of Medicine, University of Crete, Heraklion.
Hellenic Virology, 1996, 1(1): 51-55.
To determine the significance of sludge and shellfish as vehicles for enteric viruses transmission to men, convinient and reliable methods including extraction from sludge and shellfish tissues are needed to detect these viruses. The development of an extraction/concentrationprocedure to recover Hepatitis A virus (HAV) from wastewaters and sludge and oyster carcasses is the purpose of this study. Homogenates of oyster tissues or sludges were inoculated with cell culture adapted HAV (strain CF53) and the virus was extracted by elution with 3% beef extracts at pH 9,5 followed by a clarification of the eluate by centrifugation. Then, HAV from eluate supernatant was concentrated by polyethylene glycol (PEG 6000) precipitation. The obtained precipitate ws resuspended in the use of eluent, clarified by centrifugation reconcentrated by PEG 6000 precipitation. Hepatitis A antigen was detected by a solid - phase radioimmunoassay and quantified by RIA-endpoint titration. Infectious HAV was quantified by cell culture titration by using PLC/PRF/5 cell line. Results showed that HAV can be efficiently recovered from sludge or oyster titers by elution and twofold PEG 6000 precipitation, with a mean average recovery greater than 65% of the initial virus added to the samples. By using this procedure, wild strains of HAV were detected in several samples coming from three European countries after regular shellfish or sludge collection. Thus, we recommend the use of the described elution/precipitation method for detecting pollution with viral hepatitis in samples recommend the use of the described elution/precipitation method for detecting pollution with viral hepatitis in samples of molluscan shellfishes or wastewater sludge.
Prevalence of hepatitis viruses A in rural and urban population in a region of Crete.
M. Tsopanomichalou (1), N. Fragkiadakis (2), A. Karachristos (1), E. Biziagos (1).
(1) Clinical Virology Laboratory and (2) Gastroenterology Clinic, Department of Medicine, University of Crete, Heraklion.
Hellenic Virology, 1996, 1(1): 56-59.
Viral hepatitis is a serious problem in the island of Crete. The prevelance of hepatitis viruses in rural and urban population in a region of Crete was studied. Anti-HAV, HBsAg, anti-HBc, anti-HCV, anti-HDV and anti-HEV were determined by enzyme immunoassay in 361 healthy individuals (> 18 years old) from four rural and one urban areas of Crete. Anti-HCV, anti-HDV and anti-HEV antibodies were detected respectively in 92.0, 1.3 and 6.7% of individuals from the rural area and in 87,5, 4,4 and 5,9% of individuals from the urban area. HBsAg was found in 0,4 and 1,5% of the examined persons from the urban and rural area respectively, while, anti-HBc was detected in 25,8 and 30,9 of the examined persons respectively. Anti-HDV was not detected in all HBsAg positive samples examined.
Immunity to poliomyelitis of the Northern Greek population 30 years after the introduction of the Sabin vaccine.
V. Kyriazopoulou-Dalaina, F. Frantzidou-Adamopoulou, E. Diza-Mataftsi, E. Souliou-Symeonidou.
Microbiology Laboratory, Aristotelian University of Thessaloniki
Hellenic Virology, 1996, 1(1): 60-64.
In order to investigate the immunity levels of the Northern Greek population to Polioviruses, 30 years after the mass introduction of the Sabin vaccine, 614 sera from healthy people aged 1 day to 75 years old were examined. The method used was the micrometabolic inhibition test for the estimation of Poliovirus neutralizing antibodies (Kyriazopoulou and Bell, 1972). According to the results 80.1%, 80.2% and 84.6% of the people examine had antibodies at protective levels (>1:8) to Polioviruses I, II and III respectively. Full protection was found in 66.6% (antibodies to all types of Poliovirus) while 3% lacked antibodies to any of the Polioviruses. Referring to age groups, neonates 1-90 days old, had high levels of antibodies at rates of 90.9%, 92.4% and 90.9% for Polioviruses I, II and III respectively. The antibody levels for the rest age groups ranged from 72.4% to 86.4% for Polio I, from 68.4% to 86.4% for Polio II and from 81.5% to 89.2% for Polio III. The lower rates were found in old people >65 years old. The usual antibody titres were from 32 to 128. These results are compared with analogous results in other countries and with results of the same region found ten years ago (1984).
Papillomaviruses: Introduction, Classifiction, Physicochemical properties and Genomic Structure.
A. Tsotsos
Microbiology Department, Athens University Medical School.
Hellenic Virology, 1996, 1(1): 67-69.
The family Papovaviridae consists of two genera, Papillomavirus and Polyomavirus. Papovaviruses are small, non-enveloped, icosahedral virions with double-stranded, circular DNA. The two genera differ in the size of the virion, the m.w.of the DNA and their biological properties. Papovaviruses of both genera cause chronic and latent infections in humans and a variety of animal species. Human papillomaviruses (HPV) infect cutaneous and mucosal epithelial cells. Special attention has been drawn to HPV, as it was realized that, at least some genotypes, are closely associated with tumour formation and specifically with cervical carcinoma. Regions of HPV DNA that encode viral proteins are located on one strand (the plus strand) and are divided into th early and late regions. Early proteins E1 to E8 are functional proteins, important in transcription, replication and oncogenic transformation. Late proteins L1 and L2 are structural proteins. Papillomaviruses do not grow in conventional cell cultures. They require specific nuclear, transcriptional factors for their growth, and for this reason they can multiply only in differentiating stratified squamous epithelium, which releases these factors. This need of papillomaviruses explains, to a certain extent, the limited tropism of papillomavisuses for certain animal species and tissues. The E6 and E8 proteins of HPV 16 and 18 bind and inactivate, respectively, the cellular growth (transformation) suppressor proteins p53 and p105 RB (retinoblastoma gene product).
Histopathology of human papilloma virus (HPV) lesions and techniques for HPV detection.
G. Kontogeorgos.
Anatomic Pathology Laboratory, Peripheral General Hospital of Athens
Hellenic Virology, 1996, 1(1): 72-76.
Koilocytosis represents a common feature of HPV related papillary lesions of the squamous epithelium. In addition, clumps of keratohyaline are present in common warts. Among HPV lesions, flat changes of the lower genital track are of particular interest. The iimmunohistochemical localisation of HPV is based on the demonstration of capsid proteins by specific antibody The in situ hybridisation is a useful tool for the detection and typing of HPV, within intact cells with preserved morphology. The PCR-in situ combines the amplification of HPV genome with the demonstration of the product by conventional in situ hybridization.
Animal papillomaviruses.
M. Koumbati-Artopiou.
Veterinary Department, Aristotelian University of Thessaloniki
Hellenic Virology, 1996, 1(1): 79-82.
Animal papillomaviruses affect many species of domesticated or wild mammals and also some species of birds. Bovine are infected more frequently than equine, dogs, rabbits and sheep. Papillomaviruses have also detected in different species of beer and also from coyots, monkeys, antelopes, elephants, and panthers. Papillomas or fibropapillomas are induced either on the body skin or on mucosal membrane of digestive tract. They are usually begining and self-limitted. However, in some cases papillomas have been observed to be contiguous with a squannous cell carcinoma, especially when other factors like exposure to sunlight, ingestion of braken fern or lack of skin melanine pigments have an influence at the same time.
The polymerase chain reaction (PCR) in the diagnosis of viral infections.
P. Markoulatos (1), V. Samara (1), G. Marinakis (1), K. Kaparos (1), P. Foutoucidou (1), V. Krikelis (1), N. Spyrou (1), N. Vamvakopoulos (2).
(1) Virology Department, Hellenic Pasteur Institute, Vas. Sofias Ave. 127, Athens 115 21 and (2) Laboratory of Biology & Genetics, Department of Medicine, University of Thessaly.
Hellenic Virology, 1996, 1(1): 83-88.
We outline the basic principles and key features, including specificity and sensitivity of PCR and comment on its high risk of contamination, considered to be main cause of false positivity in "Nested PCR" applications. A broad spectrum of diagnostic virological PCR applications including Reverse Transcription PCR, for detection and typing of viruses HIV 1 & 2, HCV, HSV 1 & 2,CMV, EBV, and Polioviruses 1,2,3, vaccinal or wild strains is also being discussed.
Comparison of indirect immunofluorescence for the serological diagnosis of EBV infections (VCA, EA, EBNA) and ELISA using EBV polypeptides from recombinant DNA technology.
N. Spyrou, P. Markoulatos, M. Kapsali, B. Krikelis, A. Mentis.
Virology Department, Hellenic Pasteur Institute, Vas. Sofias Ave. 127, Athens 115 21.
Hellenic Virology, 1996, 1(1): 89-92.
The micro-ELISA procedure of Biotest using EBV polypeptides (EAD p54, p138 and EBNA p-72) from recombinant DNA technology was compared to immunofluorescence assay of Gull Laboratories for the diagnosis and determination of clinical status of EBV infected patients. In primary infections the sensitivity was 62,5% and specificity 100%. The micro-ELISA procedure of Biotest can be succesfully for the diagnosis of primary EBV infections while in the case of viral reactivation serial serum samples are necessary in order to determine with precission the antibody cinetique to different EBV induced antigens(VCA IgG and IgM, EA IgG and IGM and EBNA IgG).