95-11294. Viruses, Serums, Toxins, and Analogous Products; Detection of Extraneous Agents by the Fluorescent Antibody Technique  

  • [Federal Register Volume 60, Number 89 (Tuesday, May 9, 1995)]
    [Rules and Regulations]
    [Pages 24547-24549]
    From the Federal Register Online via the Government Publishing Office [www.gpo.gov]
    [FR Doc No: 95-11294]
    
    
    
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    DEPARTMENT OF AGRICULTURE
    9 CFR Part 113
    
    [Docket No. 93-071-2]
    
    
    Viruses, Serums, Toxins, and Analogous Products; Detection of 
    Extraneous Agents by the Fluorescent Antibody Technique
    
    AGENCY: Animal and Plant Health Inspection Service, USDA.
    
    ACTION: Final rule.
    
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    SUMMARY: We are amending the regulations concerning testing by the 
    fluorescent antibody technique for extraneous agents (viruses) in cells 
    of animal origin that are used in the manufacture of veterinary 
    biologics. The amendment allows the use of alternative fluorochromes 
    that may be conjugated to an antibody, revises the list of extraneous 
    agents to be tested for, and includes extraneous agents for which 
    equine cells are to be tested. In addition, the word ``agent'' is 
    replaced with the word ``virus'' since this is the agent being tested 
    for. The amendment is necessary to update the requirements related to 
    the testing for extraneous viruses.
    
    EFFECTIVE DATE: June 8, 1995.
    
    FOR FURTHER INFORMATION CONTACT:
    Dr. David A. Espeseth, Deputy Director, Veterinary Biologics, BBEP, 
    APHIS, 4700 River Road Unit 148, Riverdale, MD 20737-1237, (301) 734-
    8245.
    
    SUPPLEMENTARY INFORMATION: 
    
    Background
    
        In accordance with the regulations contained in 9 CFR part 113, 
    standard requirements are prescribed for the preparation of veterinary 
    biological products. A standard requirement consists of specifications, 
    procedures, and test methods which define the standards of purity, 
    safety, potency, and efficacy for a given type of veterinary biological 
    product. Microorganisms, animal cells, and ingredients of animal origin 
    used in production are required to be tested for extraneous viruses. In 
    part, this involves testing for the presence of extraneous viruses by 
    the fluorescent antibody technique described in Sec. 113.47. When the 
    current standard requirement was established, fluorescent antibodies 
    were constructed by conjugating antibodies to one of the fluorochromes, 
    fluorescein. Fluorochromes are any of a variety of chemicals used in 
    cytochemistry to produce a secondary fluorescence in the specimen. In 
    the intervening years, additional fluorochromes have been developed for 
    use as cytochemical markers or stains.
        Standard requirements included in the regulations specify that 
    cells, master seed virus, and most ingredients of animal origin used in 
    the production of biological products be tested for contaminating 
    bacteria, fungi, mycoplasma, cytopathogenic organisms, viruses, 
    hemadsorbing agents, and extraneous agents (viruses) detectable by the 
    fluorescent antibody technique. The presence of specific fluorescence 
    associated with the use of certain antibodies, in comparison with the 
    [[Page 24548]] appropriate controls, is an indication of the presence 
    of the contaminating antigen or extraneous virus against which the 
    antibody was made.
        Current Sec. 113.47 lists the types of extraneous viruses against 
    which fluorescein-conjugated antibodies are to be used in testing cells 
    from certain species of animals. New viruses have since been identified 
    as animal pathogens. No viruses which are disease agents of horses are 
    included in the current Sec. 113.47. As new knowledge has developed, 
    testing for these agents has become necessary.
        On March 21, 1994, we published in the Federal Register (59 FR 
    13257-13259, Docket No. 93-071-1) a proposal to amend the regulations 
    by revising Sec. 113.47 to allow the use of additional fluorescent 
    stains in the testing of extraneous viruses by the fluorescent antibody 
    technique. The current regulations in Sec. 113.47 limit the stain used 
    in the fluorescent antibody test to fluorescein (a fluorochrome). Other 
    fluorochromes, when conjugated to antibodies, may be expected to 
    perform as well as fluorescein in the test for extraneous viruses. The 
    amendment thus allows for the use of alternative fluorochromes in such 
    tests. The term ``fluorescein-conjugated antibody'' is replaced with 
    ``fluorochrome-conjugated antibody'' everywhere it appears in 
    Secs. 113.47 and 113.52(b)(2) (i) and (ii).
        The current regulations in Sec. 113.47 lists the specific 
    extraneous viruses against which antibodies are used in the testing of 
    certain types of cells. We have revised the list of cell types to be 
    tested for extraneous viruses to include equine cells. Those using 
    other cells for the production of biologics may also be required to 
    test for specific viruses before such use is approved.
        We solicited comments concerning our proposal for a 60-day comment 
    period ending April 20, 1994. We received one comment by that date from 
    a manufacturer of veterinary biological products. We carefully 
    considered the comment which is discussed below.
        The comment was in two parts The first was that the fluorescent 
    antibody (FA) test was not as sensitive for detecting bluetongue virus 
    as seroconversion in sheep, and the use of the FA test would require 
    firms to introduce this organism into their testing facility. The 
    second was that testing of bovine cells for bovine respiratory 
    syncytial virus was also a new requirement.
        In response to the comment, the Animal and Plant Health Inspection 
    Service has found that the FA test is a sensitive, specific, and 
    accurate test for the detection of extraneous viruses in seeds, cells 
    and ingredients of animal origin. When used with the required controls, 
    the FA test detects the presence of specific antigens indicative of the 
    presence of specific extraneous viruses. The use of positive and 
    negative controls with the FA test eliminates the incidence of false 
    positives and false negatives. The FA test also has the advantage of 
    relieving the firms from the need to locate and maintain seronegative 
    sheep.
        APHIS acknowledges that requiring firms to test for the bluetongue 
    virus will result in the introduction of the agent onto their premises. 
    This should, however, pose no greater problem to the firms than 
    introducing other viruses for the purpose of conducting these tests. If 
    no firm's biosecurity is adequate to contain such viruses as those 
    causing bovine virus diarrhea, canine parvoviral diarrhea, or 
    pseudorabies, it should be able to contain bluetongue virus. Further, 
    there is no threat of human disease from the use of bluetongue virus in 
    association with this test as bluetongue virus is not known to affect 
    humans.
        With respect to the requirement for testing bovine cells for bovine 
    respiratory syncytical virus, APHIS has determined that this virus may 
    be present as an extraneous agent in cells of bovine origin. One of the 
    major purposes of the rule is to update the list of extraneous viruses 
    for which seeds, cells, and ingredients of animal origin are tested. 
    The new requirements are based on current knowledge of animal diseases. 
    No changes are made to the regulations in response to the commenter.
        Therefore, based on the rationale set forth in the proposed rule 
    and in this document, we are adopting the provisions of the proposal as 
    a final rule with one conforming change in Sec. 113.300(c)(1).
    
    Executive Order 12866 and Regulatory Flexibility Act
    
        This final rule has been reviewed under Executive Order 12866. This 
    rule has been determined to be not significant for the purposes of 
    Executive Order 12866, and, therefore, has not been reviewed by the 
    Office of Management and Budget.
        These amendments should not have a significant economic impact on 
    manufacturers since they will broaden the range of fluorochrome stains 
    that may be used in conducting the fluorescent antibody test and will 
    revise the list of extraneous agents for which various cell types are 
    to be tested with the fluorescent antibody technique. The amendments 
    will thus remove outdated requirements and provide flexibility in the 
    types of antibody that may be used in tests for extraneous agents. On 
    balance, therefore, there should be no net increase in testing 
    requirements over the current requirements.
        Under these circumstances, the Administrator of the Animal and 
    Plant Health Inspection Service has determined that this action will 
    not have a significant economic impact on a substantial number of small 
    entities.
    
    Executive Order 12372
    
        This program/activity is listed in the Catalog of Federal Domestic 
    Assistance under No. 10.025 and is subject to Executive Order 12372, 
    which requires intergovernmental consultation with State and local 
    officials. (See 7 CFR part 3015, subpart V.)
    
    Executive Order 12778
    
        This final rule has been reviewed under Executive Order 12778, 
    Civil Justice Reform. This rule: (1) Preempts all State and local laws 
    and regulations that are inconsistent with this rule; (2) has no 
    retroactive effect; and (3) does not require administrative proceedings 
    before parties may file suit in court challenging this rule.
    
    Paperwork Reduction Act
    
        This rule contains no information collection or recordkeeping 
    requirements under the Paperwork Reduction Act of 1980 (44 U.S.C. 3501 
    et seq.).
    
    List of Subjects in 9 CFR Part 113
    
        Animal biologics, Exports, Imports, and Reporting and recordkeeping 
    requirements.
    
        Accordingly, 9 CFR part 113 is amended as follows:
    
    PART 113--STANDARD REQUIREMENTS
    
        1. The authority citation for part 113 continues to read as 
    follows:
    
        Authority: 21 U.S.C. 151-159; 7 CFR 2.17, 2.51, and 371.2(d).
    
        2. Section 113.47 is revised to read as follows:
    
    
    Sec. 113.47  Detection of extraneous viruses by the fluorescent 
    antibody technique.
    
        The test for detection of extraneous viruses by the fluorescent 
    antibody technique provided in this section shall be conducted when 
    prescribed in an applicable Standard Requirement or in a filed Outline 
    of Production for a product.
        (a) Monolayer cultures of cells (monolayers), at least 7 days after 
    the [[Page 24549]] last subculturing, shall be processed and stained 
    with the appropriate antiviral fluorochrome-conjugated antibody as 
    specified in paragraph (b) of this section.
        (1) Three groups of one or more monolayers shall be required for 
    each specific virus prescribed in paragraph (b) of this section.
        (i) At the time of the last subculturing, one group of test 
    monolayers shall be inoculated with approximately 100-300 FAID50 
    of the specific virus being tested for as positive controls.
        (ii) One group of monolayers shall be the ``material under test.''
        (iii) One group of monolayers, that are of the same type of cells 
    as the test monolayers and that have been tested as prescribed in 
    Secs. 113.51 or 113.52 (whichever is applicable), shall be prepared as 
    negative controls.
        (2) Each group of monolayers shall have a total area of at least 6 
    cm\2\.
        (3) Positive control monolayers may be fixed (processed so as to 
    arrest growth and assure attachment of the monolayer to the surface of 
    the vessel in which they are grown) before 7 days after subculturing if 
    fluorescence is enhanced by doing so, Provided, That a monolayer of the 
    material under test is also fixed at the same time as the positive 
    control and a monolayer of the material under test is also fixed at 
    least seven days after subculturing. Monolayers that are fixed before 7 
    days after subculturing shall be stained at the same time as the test 
    monolayers and negative controls fixed at least 7 days after 
    subculturing.
        (b) The antiviral fluorochrome-conjugated antibodies to be used 
    shall depend on the type of cells required to be tested for extraneous 
    viruses as specified in an applicable Standard Requirement or in a 
    filed Outline of Production. Antiviral fluorochrome-conjugated 
    antibodies specific for the extraneous viruses shall be applied to each 
    respective type of cell in accordance with the following list. Under 
    certain circumstances, additional tests may need to be conducted, as 
    determined by the Administrator. When a specific antiviral 
    fluorochrome-conjugated antibody is used in testing for the listed 
    extraneous viruses specified in more than one cell type, it need only 
    be applied to the most susceptible cell type.
    
    (1) All cells shall be tested for:
        (i) Bovine virus diarrhea virsus;
        (ii) Reovirus; and
        (iii) Rabies virus.
    (2) Bovine, caprine, and ovine cells shall, in addition, be tested for:
        (i) Bluetongue virus;
        (ii) Bovine adenoviruses;
        (iii) Bovine parvovirus; and
        (iv) Bovine respiratory syncytial virus.
    (3) Canine cells shall, in addition, be tested for:
        (i) Canine coronavirus;
        (ii) Canine distemper virus; and
        (iii) Canine parvovirus.
    (4) Equine cells shall, in addition, be tested for:
        (i) Equine herpesvirus; and
        (ii) Equine viral arteritis virus.
    (5) Feline cells shall, in addition, be tested for:
        (i) Feline infectious peritonitis virus; and
        (ii) Feline panleukopenia virus.
    (6) Porcine cells shall, in addition, be tested for:
        (i) Porcine adenovirus;
        (ii) Porcine parvovirus;
        (iii) transmissible gastroenteritis virus; and
        (iv) Porcine hemagglutinating encephalitis virus.
    (7) Firms that do not have rabies virus on premises either for research 
    or production purposes are exempt from having to produce positive 
    rabies virus control monolayers. Fixed positive rabies virus control 
    monolayers will be provided by the National Veterinary Services 
    Laboratories.
    
        (c) After staining, each group of monolayers shall be examined for 
    the presence of specific fluorescence attributable to the presence of 
    extraneous viruses.
        (1) If the material under test shows any evidence of specific viral 
    fluorescence, it is unsatisfactory and may not be used; Provided, That, 
    if specific fluorescence attributable to the virus being tested for is 
    absent in the positive control monolayers, the test is inconclusive and 
    may be repeated.
        (2) If the fluorescence of the monolayers inoculated with the 
    specific virus as positive controls is equivocal, or if the negative 
    monolayers show equivocal fluorescence indicating possible viral 
    contamination, or both, the test shall be declared inconclusive, and 
    may be repeated; Provided, That, if the test is not repeated, the 
    material under test shall be regarded as unsatisfactory for use in the 
    production of biologics.
        3. Section 113.52, paragraph (b)(2)(i) and (ii), is revised to read 
    as follows:
    
    
    Sec. 113.52  Requirements for cell lines used for production of 
    biologics.
    
    * * * * *
        (b) * * *
        (2) * * *
        (i) At least two monolayers shall be stained with an antispecies 
    fluorochrome-conjugated antibody unrelated to the species of origin of 
    the MCS.
        (ii) At least two monolayers shall be stained with an antispecies 
    fluorchrome-conjugated antibody specific to the species of origin of 
    the MSC.
    * * * * *
    
    
    Secs. 113.51, 113.52 and 113.53  [Amended]
    
        4. In the following places, the word ``agents'' are removed and the 
    word ``viruses'' added in its place:
        a. Section 113.51, paragraph (c)(3)(ii).
        b. Section 113.52, paragraph (f)(4)(ii).
        c. Section 113.53, paragraph (c)(6)(ii).
        5. In Sec. 113.300, paragraph (c)(1), the term ``fluorescein'' is 
    removed and the term ``fluorochrome'' added in its place.
    
        Done in Washington, DC, this 1st day of May 1995.
    B. Glen Lee,
    Acting Administrator, Animal and Plant Health Inspection Service.
    [FR Doc. 95-11294 Filed 5-8-95; 8:45 am]
    BILLING CODE 3410-34-M
    
    

Document Information

Effective Date:
6/8/1995
Published:
05/09/1995
Department:
Agriculture Department
Entry Type:
Rule
Action:
Final rule.
Document Number:
95-11294
Dates:
June 8, 1995.
Pages:
24547-24549 (3 pages)
Docket Numbers:
Docket No. 93-071-2
PDF File:
95-11294.pdf
CFR: (2)
9 CFR 113.47
9 CFR 113.52