Instituto Valenciano de Microbiología
(IVAMI)

Masía El Romeral
Ctra. de Bétera a San Antonio Km. 0.3
46117 Bétera (Valencia)
Phone. 96 169 17 02
Fax 96 169 16 37
Email: 
www.ivami.com
CIF B-96337217

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Virucidal test against hepatitis B virus, using as surrogated virus, Herpes suino virus type 1 (Pseudorabies), following the methods recommended by the UNE-EN 14476: 2014 + A1: 2015 standard. Chemical disinfectants and antiseptics. Quantitative suspension test for the evaluation of virucidal activity in the medical area. Test method and requirements (phase 2, Step 1).

Test not accredited in our laboratory (see note 1, for accredited test).

Test with the certificate of Good Laboratory Practices (GLPs).

Note: The UNE-EN 14476 test, for general virucidal activity, requires the following three viral strains to be tested: Poliovirus type 1, Adenovirus type 5 and Murine Norovirus. This is because these viruses are more resistant to the action of disinfectants and when a disinfectant is active against them can be considered effective against other viruses not tested. If the test is performed with other species/strains (for example, in this case, hepatitis B virus), it can be certified that the test shows activity against this species/viral strain, but does not meet the requirement of the standard guideline to be considered virucidal in general.

            For the Hepatitis B virus (HBV) there is no cell line for its replication in vitro. However, the use of substitute viruses with similar biophysical properties and genomic structure allows the evaluation of the virucidal efficacy of chemical disinfectants in quantitative suspension tests. The recommended surrogate virus is the swine herpes virus type 1 (Pseudorabies) that can be replicated in cell cultures in vitro. Another of the most recommended substitution viruses is duck viral hepatitis, which replicates in primary cultures of duck embryo liver (DEL) cells. However, given the sensitivity and difficulty in obtaining these cells, in our laboratory and in several publications (among them, WHO Technical Report, Series No. 924, 2004), the swine herpes virus type 1 (Pseudorabies) is used. The method used is the same as indicated in the guide EN 14476.

            In addition, the new version UNE-EN 14476: 2014 + A1: 2015, introduces the possibility that the Vaccinia virus can be used when testing disinfectants against enveloped viruses for hygienic handwashing and hygienic handrub. Therefore, as the hepatitis B virus (HBV) is a enveloped virus, we offer the test for the activity of disinfectants against the hepatitis B virus using the herpes virus type 1 (Pseudorabies), or with the virus Vaccinia, when disinfectants are tested for hygienic handwashing or hygienic handrub.

            The test conditions are the following: 1) Hygienic hands treatment by handrub and hygienic handwashing, when a limited spectrum is desired - covered viruses-; test temperature 20°C; contact time between 30 and 120 seconds*; and clean conditions for the hygienic treatment by handrub, or dirty for handwashing; 2) Disinfection of instruments 20ºC test temperature; contact time not exceeding 60 min*; clean or dirty conditions *; 3) Disinfection of surfaces, test temperature between 4 and 30ºC*; maximum contact time of 5 minutes, for surfaces close to patients or 60 minutes maximum for other surfaces*; clean or dirty conditions*. The product should be tested at least three concentrations including one in the active range and one in the non-active range (these last two concentrations are chosen by the laboratory that performs the test since they correspond to the positive and negative controls, respectively). To consider the product as virucidal, a logarithmic reduction of 4 in infectivity must be demonstrated (*: the applicant for the test must choose within the ranges or alternatives indicated, and include it in the test form).

Note 1: In order for the activity against optional viruses to be covered by the ENAC accreditation, the tests must also be carried out with one of the viruses required by the standard (Poliovirus type 1, Adenovirus type 5 or murine Norovirus).