[Federal Register Volume 59, Number 54 (Monday, March 21, 1994)]
[Unknown Section]
[Page 0]
From the Federal Register Online via the Government Publishing Office [www.gpo.gov]
[FR Doc No: 94-6550]
[[Page Unknown]]
[Federal Register: March 21, 1994]
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DEPARTMENT OF AGRICULTURE
9 CFR Part 113
[Docket No. 93-071-1]
Viruses, Serums, Toxins, and Analogous Products; Detection of
Extraneous Agents by the Fluorescent Antibody Technique
AGENCY: Animal and Plant Health Inspection Service, USDA.
ACTION: Proposed rule.
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SUMMARY: We are proposing to amend 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 proposed amendment would allow the use of
alternative fluorescent-antibody conjugated antibodies; revised the
list of extraneous agents to be tested for; and include extraneous
agents for which equine cells are to be tested. The proposed amendment
is necessary to update the requirements related to the testing for
extraneous viruses.
DATES: Consideration will be given only to comments received on or
before April 20, 1994.
ADDRESSES: Please send an original and three copies of your comments to
Chief, Regulatory Analysis and Development, PPD, APHIS, USDA, room 804,
Federal Building, 6505 Belcrest Road, Hyattsville, MD 20782. Please
state that your comments refer to Docket No. 93-071-1. Comments
received may be inspected at USDA, room 1141, South Building, 14th
Street and Independence Avenue SW., Washington, DC, between 8 a.m. and
4:30 p.m., Monday through Friday, except holidays. Persons wishing to
inspect comments are requested to call ahead on (202) 690-2817 to
facilitate entry into the comment reading room.
FOR FURTHER INFORMATION CONTACT:
Dr. David A. Espeseth, Deputy Director, Veterinary Biologics, BBEP,
APHIS, USDA, room 838, Federal Building, 6505 Belcrest Road,
Hyattsville, MD 20782, (301) 436-8245.
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 by regulations in part 113 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
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 been necessary.
Changes and Clarifications
The title of Sec. 113.47 refers to the detection of extraneous
agents. Since the section is specific to viruses, the title would be
amended accordingly.
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. This
proposed amendment would allow the use of any fluorochrome-conjugated
antibody in such tests. The term ``fluorescein-conjugated antibody''
would be replaced with ``fluorochrome-conjugated antibody'' everywhere
it appears in Sec. 113.47 and in Sec. 113.52(b)(2) (i) and (ii).
The current regulation in Sec. 113.47(a) provides for staining cell
monolayers for parvovirus as early as 4 days after subculturing. Other
viruses may also be detected by processing at less than 7 days.
Therefore, the specific exemption for parvovirus has been deleted in
Sec. 113.47(a), and the provision in Sec. 113.47(a)(3) for fixing
control monolayers at less than 7 days has been modified to require
that a test monolayer also 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) at the same time. In addition, another monolayer
of the material under test and the negative control monolayer must be
held for at least 7 days before fixing. Further clarification is made
to Sec. 113.47(a)(3) that all monolayers would be stained concurrently,
regardless of time of fixing, at least 7 days after subculture.
The current regulations in Sec. 113.47 lists the specific
extraneous viruses against which antibodies are to be used in the
testing of certain types of cells. The cells with which the fluorescent
antibody test is to be used are specified in Secs. 113.51, 113.52,
113.53, or 113.55. In the proposed amendment to Sec. 113.47, Vero cells
would not be listed as one of the specific cell types since they are
specified in those sections as listed above. We have also 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 specified viruses before such use is
approved.
Proposed amendments to Sec. 113.47(b) pertain to the extraneous
viruses associated with specific cells that would be tested for using
specific fluorochrome-conjugated antibodies. All cells (including
caprine and equine would be tested for bovine virus diarrhea virus,
reovirus, and rabies virus. In addition, bluetongue virus would be
added to the list of extraneous agents for testing bovine cells, canine
coronavirus would be listed for testing canine cells, feline infectious
peritonitis virus would be specified in place of coronavirus for
testing feline cells, and porcine hemagglutinating encephalitis virus
would be added for testing porcine cells. Equine cells would be tested
with fluorochrome-conjugated antibodies to equine herpesvirus and to
equine viral arteritis virus.
In addition, non-substantive minor editorial changes to the
regulations are made for clarity.
Executive Order 12866 and Regulatory Flexibility Act
This rule has been determined to be not significant for purpose of
Executive Order 12866 and therefore has not been reviewed by OMB.
These proposed amendments should not have a significant economic
impact on manufacturers since it would broaden the range of
fluorochrome-labeled antibodies that may be used in conducting the
fluorescent antibody test and would revise the list of extraneous
agents for which various cell types are to be tested with the
fluorescent antibody technique. The proposed amendments would thus
remove outdated requirements and provide flexibility in the types of
antibody that may be used in tests for extraneous agents.
Under these circumstances, the Administrator of the Animal and
Plant Health Inspection Service has determined that this action would
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 proposed rule has been reviewed under Executive Order 12778,
Civil Justice Reform. If this proposed rule is adopted: (1) All State
and local laws and regulations that are in conflict with this rule will
be preempted; (2) no retroactive effect will be given to this rule; and
(3) administrative proceedings will not be required before parties may
file suit in court challenging this rule.
Paperwork Reduction Act
This proposed rule contains no information collection or
recordkeeping requirements under the Paperwork Act of 1980 (44 U.S.C.
3501 et seq.)
List of Subjects in 9 CFR Part 113
Animal biologics, Exports, Imports, Reporting and recordkeeping
requirements.
Accordingly, 9 CFR part 113 would be amended as follows:
PART 113--STANDARD REQUIREMENTS
1. The authority citation for part 113 would continue 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 would be 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 last subculturing, shall be processed and stained with the
appropriate antiviral fluorochorome-conjugated antibody as specified in
paragraph (b) of this action.
(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
cm2.
(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 virus;
(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 calles 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), would be 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
fluorochrome-conjugated antibody specific to the species of origin of
the MCS.
* * * * *
Secs. 113.51, 113.52, and 113.53 [Amended]
4. In the following places, the word ``agents'' would be 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).
Done in Washington, DC, this 15th day of March 1994.
Patricia Jensen,
Acting Assistant Secretary, Marketing and Inspection Services.
[FR Doc. 94-6550 Filed 3-18-94; 8:45 am]
BILLING CODE 3410-34-M