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At least 19 recordsLinked to original sources
PubMedDate not supplied

Avian cell cultures grown in an open system (dishes) without CO2 incubator, and their use for virus cultivation.

An organic buffer, N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES), in a culture medium made it possible to culture chick embryo fibroblast (CEF), duck embryo fibroblast (DEF), and chick kidney (CK) cells in an open system (dishes) under incubation in a bacteriological incubator, requiring no CO2 incubation. CEF, DEF, and CK cells cultured with this system were suitable for cultivation of avain viruses such as Newcastle disease virus, CELO virus, Marek's disease virus, and turkey herpesvirus.

Adenoviridae↗
PubMed1994

Use of PCR-enzyme immunoassay for identification of influenza A virus matrix RNA in clinical samples negative for cultivable virus.

Influenza A virus infections are a major cause of morbidity and mortality worldwide. Standard diagnostic methods either are not efficient in identifying infected individuals in a timely manner or lack sensitivity. We developed a PCR-enzyme immunoassay (PCR-EIA) for the detection of influenza A virus RNA in respiratory secretions. A reverse transcription PCR was performed with oligonucleotide primers directed at a highly conserved area of the influenza A matrix gene. Amplified DNA was identified by hybridization in solution to a nested biotinylated RNA probe and quantitated in an EIA. PCR-EIA detected small quantities of RNA from the three prevalent subtypes of human influenza A virus. Influenza B and C, parainfluenza, measles, mumps, and respiratory syncytial viruses tested negative. The potential efficiency of PCR-EIA for use in clinical diagnosis was determined by testing 90 nasal wash specimens obtained daily over a 10-day period from nine human volunteers infected with influenza A virus. Thirty-seven of the postinfection samples had detectable influenza A virus RNA by PCR-EIA, whereas only 26 postinfection samples were positive by culture. PCR-EIA was particularly efficient for the identification of influenza A virus in samples obtained more than 4 days after infection. Seventeen of 45 such samples were positive, whereas virus was cultivated from 4 samples (P < 0.00005). All preinfection samples from volunteers subsequently infected with influenza A virus were negative by PCR-EIA, as were samples from a volunteer infected with parainfluenza virus type 3. Nucleic acid amplification techniques represent important tools for the timely and sensitive diagnosis of influenza A virus infections and, therefore, their management and control.

Base Sequence↗
PubMed1990

Detection of human immunodeficiency virus-1 by polymerase chain reaction and virus cultivation.

Peripheral blood of 57 patients with antibodies to human immunodeficiency virus 1 (HIV-1) and of five HIV-1 seronegative subjects at risk for HIV-1 infection were analysed by polymerase chain reaction (PCR) and virus isolation. The virus was recovered from peripheral blood cells in 89% and from plasma in 75% of the HIV-1 seropositive cases. In contrast, proviral HIV-1 DNA was detected in all HIV-1 seropositive patients by dot blot hybridization of the amplified fragments. The intensities of the dot blot reactions were less pronounced in asymptomatic HIV-1 seropositive individuals than in patients with acquired immunodeficiency syndrome (AIDS) or AIDS-related complex (ARC), suggesting an increase in proviral DNA with advancing disease. Three of five seronegative patients with signs or symptoms suggesting HIV-1 infection, but none of the controls, were positive for HIV-1 DNA by one or two primer pairs. These results show a high sensitivity of the PCR for detecting HIV-1 DNA in patients of all stages of HIV-1 infection. Proviral DNA can also be detected in some individuals without detectable antibodies to the virus. The virus load in peripheral blood, as determined by virus cultivation and PCR, seems to increase with progression of the infection.

Adult↗
PubMed1987

Serological studies with influenza A(H1N1) viruses cultivated in eggs or in a canine kidney cell line (MDCK).

Pairs of influenza A(H1N1) viruses cultivated from the same clinical specimen in canine kidney (MDCK) cells or in embryonated hens' eggs can frequently be distinguished by their reactions with monoclonal antibodies to haemagglutinin and with antibodies in ferret or human sera. Egg-adapted virus, further passaged in MDCK cultures remained "egg-like" in serological characteristics indicating that the differences in their serological reactions were not a direct result of host cell-dependent glycosylation of the haemagglutinin. Haemagglutination-inhibiting (HI) or virus neutralizing antibodies in human sera can be detected more frequently, and to higher titre, in tests employing virus grown exclusively in MDCK cells than in tests with virus adapted to growth in embryonated eggs. Striking differences were detected in the serological reactions in HI tests when sera from ferrets infected with egg-grown virus were tested against a series of strains of influenza A(H1N1) virus isolated in 1983 and adapted to growth in eggs. In contrast, sera from ferrets infected with MDCK-derived virus failed to distinguish serologically between the same viruses that had been passaged exclusively in MDCK cells and also revealed relatively small differences between their egg-adapted counterparts.It was concluded that the cell substrate used for virus isolation and cultivation is a factor that should be considered when interpreting the results of strain characterization of influenza A(H1N1) isolates and in sero-surveys using these viruses.

Influenza A Virus, H1N1 Subtype↗
PubMedDate not supplied

Immunoelectrophoretic study of the host antigens of Sendai virus cultivated in different substrates (embryonated hen eggs and KB cells). Note I. Analysis of chorioallantoic membrane host antigens.

The immunoelectrophhoretic analysis of two preparations of the same virus cultivated in different substrates (Sendai/egg and Sendai/KB) demonstrate that virus envelopes contain: a) identical virus-specific antigens (but also some virus antigens with only partial immunochemical identity, or even non-identity); b) antigens incorporated by the virus from the host cell, during assembly and release. The latter differ according to the host, and some of them are only partially identical with the corresponding antigens of the host cell, which suggests that host cell glycoproteins are incorporated into the virus envelope after certain virus-induced modifications.

Animals↗
PubMed1976

[Allergic reactions in cattle vaccinated against foot-and-mouth disease with vaccines prepared from viruses cultivated in BHK cells in roller bottles].

Tests on bovines vaccinated 3 or 4 times against foot-and-mouth disease with vaccines based on viruses cultivated on BKH cells, in roller bottles, have allowed to define the nature of some of the principal allergens: on the one hand residuary horse serum used to saturate the filters employed in preparing the virus intended for the manufacture of vaccines and, on the other hand, the substances liberated by the cytopathogenic action of the virus on the BHK cells. The horse serum is the most active and causes general and local reactions which are sometimes severe. It is therefore replaced by calf serum intended to reduce the importance of the problem for bovines. BHK antigens (BHK extracts and virus) produce only local reactions which are generally less severe.

Animals↗
PubMedDate not supplied

Immunoelectrophoretic study on the host antigens of Sendai virus cultivated in different substrates (embryonated hen eggs and KB cells). Note II. Analysis of KB cell host antigens.

It could be demonstrated by immunoelectrophoretic analysis that the host antigen incorporated in the envelopes of Sendai virus cultivated in KB cells (Sendai/KB) is characteristic of these cells and does not occur in the envelopes of the same virus grown in the embryonated hen egg (Sendai/egg). This host antigen appears from the first passage in KB cells and is constantly maintained over subsequent passages. The glycoproteins released by Triton X-100 disruption from Sendai/KB envelopes have a concomitant antigenic specificity for both virus and host. In the case of Sendai/egg virus, exclusively virus-specific macromolecules are also released.

Antigens↗
PubMed1986

[Studies on canine distemper virus. I. Virus cultivation in cell cultures].

The author studied the multiplication capacity of 2 avianized strains of canine distemper virus - Onderstepoort and Lederle Encephalitis and 2 country strains - WS-66 and LL-68 in primary cell cultures of chicken embryo fibroblasts and the dog's kidney. The strain Onderstepoort multiplied and caused cytopathological changes in the cell culture of chicken embryo fibroblasts after passaging from villousallantoic membranes of chicken embryos. Maximum titer was found between the 5th and 15th passage in the cell culture and amounted to about 10(5.0) TCID50 or about 10(5.5) EID50. In all 50 passages were carried out. However, multiplication occurred, and cytopathological changes in cell culture of chicken embryo fibroblasts were caused by the strain Lederle Encephalitis only due to the application of initial adsorption and maximum inoculum by 8 successive passages. Maximum TCID50 titre occurred after 40 passages and was about 10(4.5). Both strains preserved their ability to evoke changes on villonsallantoic membranes. Studies on the effect of the inoculum size and incubation time on the virus harvest were carried out Lederle Encephalitis strain on the level of 18-20 passages. In the culture of fibroblasts the most favourable results were obtained by using 10(2.0)-10(2.5) TCID50 inoculum per 10(5.0) cells, harvested after about 96 hr of incubation. Attempts to adapt the strains Onderstepoort and Lederle Encephalitis to cell culture of the dog's kidney gave negative results. Multiplication of the country strains of canine distemper WS-66 and LL-68 was not obtained in the cell culture of the dog's kidney and in that of chicken embryo fibroblasts in the experiment conditions.

Animals↗