seeds shall be considered to mean: seeds in the botanical sense, other than those not intended for planting;

seeds, tissue cultures

plant tissue cultures.

TCID50 Tissue culture infective dose at the 50 %end point

IV.1.4. PCR confirmation of isolation of ISAV in tissue culture

(b) seedling or tissue cultures obtained in vitro, in solid or liquid media, transported in sterile containers, and

(b) seedling or tissue cultures obtained in vitro, in solid or liquid media, transported in sterile containers;

Cell cultures to be used for inoculation with tissue material should be young (4 to 48 hours old) and actively growing (not confluent) at inoculation.

For all of the following Annex A species, tissue cultures obtained in vitro, in solid or liquid media, transported in sterile containers, are not subject to the provisions of this Regulation)

Antibiotic-treated organ suspension is inoculated into cell cultures in two dilutions, i.e. the primary dilution and, in addition, a 1:10 dilution thereof, resulting in final dilutions of tissue material in cell culture medium of 1:100 and 1:1000, respectively, (in order to prevent homologous interference).

Antiserum-treated organ suspension shall be inoculated into young actively growing cell cultures to give a final dilution of tissue material to culture medium of 1:1000 For each organ suspension 40 >ISO_7>ě>ISO_1>l of inoculum shall be added to one well containing 2 ml of culture medium.

in cooperation with the suppliers of antigen and the reference laboratory for the identification of the foot-and-mouth disease virus, to review strains stored in the Community reserves and to assist in selecting suitable new strains to be adapted to tissue culture for future vaccine production;

As none of the species or higher taxa of flora included in Annex A is annotated to the effect that its hybrids shall be treated in accordance with the provisions of Article 4.1 of the Regulation, this means that artifically propagated hybrids produced from one or more of these species or taxa may be traded with a certificate of artificial propagation, and that seeds and pollen (including pollinia ), cut flowers, seedling or tissue cultures obtained in vitro, in solid or liquid media, transported in sterile containers of these hybrids are not subject to the provisions of the Regulation.

PCR confirmation of isolation of ISAV in tissue culture

If human cell or tissue cultures of specific organs are not available, other mammal cell and tissue cultures can be used.

A cell culture study should be performed in human cell or tissue cultures of different organs.

seeds, spores and pollen (including pollinia); (b) seedling or tissue cultures obtained in vitro, in solid or liquid media, transported in sterile containers, and

Negative cultures must be inoculated on fresh tissue cultures at 48 or 72 hours and this blind passage examined up to 72 hours later.

In addition, tissue culture studies suggest that cell growth in human breast tumours may be stimulated by prolactin.

Tissue culture studies suggest that cell growth in human breast tumours may be stimulated by prolactin.

seedling or tissue cultures obtained in vitro, in solid or liquid media, transported in sterile containers;

seeds and pollen; (b) seedling or tissue cultures obtained in vitro, in solid or liquid media, transported in sterile containers;

Tissue culture medium (e.g. Eagle's MEM) containing 0,04M Hepes buffer, 0,01 % bovine serum albumin and antibiotics, pH 7,2.

Prepare a cell suspension of 1 × 1055 cells/ml in culture medium and dispense 100 µl culture medium only into the peripheral wells of a 96-well tissue culture microtiter plate (= blanks).

Tissue-culture grown viral antigens are trapped by a rabbit hyperimmune anti-serum to swine vesicular disease adsorbed to the solid phase.

If undiluted serum samples are toxic to the tissue cultures, these sera may be diluted 1/2 before being used in the test. This will be equivalent to 1/4 final dilution of serum.

The test is carried out in flat-bottomed tissue culture grade microtitre plates using susceptible cells such as IB-RS-2, BHK-21 or calf kidney cells.

If the quantity and quality of samples from vesicular lesions submitted for examination is insufficient for immediate examination by ELISA, the growth of virus in tissue culture will be necessary to amplify viral antigen.

Human immunoglobulin anti-vaccinia shall contain not less than 500 IU per ml of vaccinia antibody as determined by a neutralization test on chorio-allantoic membranes or in tissue culture.

The VN test must be considered as the reference test, but has the disadvantage that it takes 2 to 3 days to complete and requires tissue culture facilities.

National Laboratories shall ensure that the tissue culture systems in use for foot-and-mouth virus isolation are sensitive to the full range of serotypes and strains for which the laboratory maintains a diagnostic capacity.

The quantitative VN micro-test for swine vesicular disease virus antibody detection is performed with IB-RS-2 cells or an equivalent cell system in flat-bottomed tissue culture grade microtitre plates.

Antiserum-treated organ suspension shall be inoculated into young actively growing cell cultures to give a final dilution of tissue material to culture medium of 1:1000 For each organ suspension 40 μl of inoculum shall be added to one well containing 2 ml of culture medium.

The simultaneous inoculation of a porcine cell line and one of the above-mentioned tissue culture systems (preferably primary bovine thyroid cells) is a useful guide as to whether vesicular samples contain swine vesicular disease virus or foot and mouth disease virus, as the former will only grow in cells of porcine origin.

Alternative monoclonal antibody-based ELISA systems, using selected monoclonal antibodies as trapping antibody and peroxidase-conjugated monoclonal antibodies as detecting antibody, may be used for swine vesicular disease antigen detection and for differential diagnosis with foot-and-mouth disease in epithelium samples, vesicular fluid or infected tissue culture.

Where sub-clinical infection is suspected, or when samples are collected after the resolution of clinical disease or when processing faecal samples, enhanced RT-PCR techniques, like nested RT-PCR, immune-PCR, ELISA-PCR and more elaborate RNA extraction methods produce a detection system at least as sensitive, but considerably more rapid, than multiple passage on tissue culture.