SENTINEL LEVEL CLINICAL
LABORATORY GUIDELINES
FOR
SUSPECTED AGENTS OF
BIOTERRORISM
AND
EMERGING INFECTIOUS DISEASES
Brucella species
American Society for Microbiology (ASM)
Revised March 2016.
For latest revision, see web site below:
https://www.asm.org/Articles/Policy/Laboratory-Response-Network-LRN-Sentinel-Level-C
Mary K. York, Ph.D.
MKY Microbiology
Consultants Walnut Creek,
ASM Subject Matter Experts: CA
ASM Sentinel Laboratory
Peter H. Gilligan, Ph.D. Protocol Working Group
University of North Carolina Hospitals/
Clinical Microbiology and Immunology Labs
Chapel Hill, NC
,Vickie Baselski, Ph.D. Barbara Robinson-Dunn, Ph.D.
University of Tennessee at Department of Clinical
Memphis Memphis, Pathology
APHL Advisory Committee
TN Beaumont Health System
Royal Oak, MI
Patricia Blevins, MPH
BRobinson-
San Antonio Metro Health District
David Craft, Ph.D.
Laboratory
Penn State Milton S. Hershey
Medical Center Michael A. Saubolle, Ph.D.
Hershey, PA Banner Health System
Erin Bowles
Phoenix, AZ
Wisconsin State Laboratory of Hygiene
Peter H. Gilligan, Ph.D. m
University of North Carolina
Hospitals/ Christopher Chadwick, MS
Susan L. Shiflett
Association of Public Health Laboratories
Clinical Microbiology and Michigan Department of
Immunology Labs
Community Health
Chapel Hill, NC Lansing, MI
Mary DeMartino, BS,
MT(ASCP)SM
State Hygienic Laboratory at the University of
Larry Gray, Ph.D. Alice Weissfeld, Ph.D.
Iowa
TriHealth Laboratories and Microbiology Specialists Inc.
University of Cincinnati College Houston, TX
of Medicine Cincinnati, OH alice@microbiologyspecialists.c Harvey Holmes, PhD
om Centers for Disease Control and
Prevention
Major Todd Kijek, Ph.D. David Welch, Ph.D.
US Army Medical Research Medical Microbiology Kara MacKeil
Institute for Infectious Diseases Consulting Association of Public Health Laboratories
Ft. Detrick, MD Dallas, TX
Chris Mangal, MPH
Michael J. Loeffelholz, Ph.D. Mary K. York, Ph.D. Association of Public Health
Department of Pathology MKY Microbiology Consultants Laboratories
Univ. Texas Medical Branch Walnut Creek, CA
Galveston, TX
Amanda Moore, BS
South Carolina Department of
Coordinating Editor: Health and Environmental Control
Judith Lovchik, Ph.D.
Indiana State Department of
James W. Snyder, Ph.D.
Health Laboratories
University of Louisville
Indianapolis, IN James Rudrik, PhD,
Louisville, KY
Michigan Department of Community
Health
Scott W. Riddell, Ph.D.
Department of Pathology Administrative Support
SUNY Upstate Medical Maureen Sullivan, MPH Minnesota
University Kimberly E. Walker, Ph.D. Department of Health
Syracuse, NY American Society for
Microbiology
2
, PREANALYTICAL CONSIDERATIONS
I. PRINCIPLE
A. Introduction
Brucella is a fastidious, aerobic, small, gram-negative coccobacillus that is neither motile
nor spore-forming (11, 19). Brucella is considered an “Overlap select agent” because it
not only has the potential to pose a threat to public health and safety, but it also poses a
threat to animal health and animal products.
This procedure describes the steps to rule out, recognize, and presumptively identify
this organism in clinical specimens in Sentinel Clinical Laboratories. Such laboratories
are defined as those certified to perform high complexity testing under the Clinical
Laboratory Improvement Amendments of 1988 (CLIA) by the Centers for Medicare
and Medicaid Services for the applicable Microbiology specialty. Laboratory in-house
testing includes Gram stains, and at least one of the following: lower respiratory tract,
wound or blood cultures.
Sentinel Clinical Laboratories are not required to register with the Select Agent
Program to conduct diagnostic testing for Select Agents, both Tier I and non-Tier 1.
Testing for Select Agents may be performed by laboratories as long as the laboratory
follows the policies listed in the reporting section of this document when a Select
Agent cannot be ruled out. Consult with your designated LRN Reference Laboratory
or refer to the CDC Division of Select Agents and Toxins website at
http://www.selectagents.gov for questions.
NOTE: To identify Brucella to the genus level some biochemical testing is needed,
however, automated systems and manual multi-test kit identifications have no place in
the identification of the organism, due to its minimal reactivity, creation of aerosols in
performance of the testing, the danger of misidentification due to its close relation to
other organisms that are positive for only a few tests, and the easy, rapid method to
presumptively identify the organism without use of system identifications. Clinically,
rapid identification to the genus level is adequate to initiate therapy, and the type of
Brucella species involved does not alter the therapy.
B. Geographic distribution
There are between 80 and 120 cases of Brucella infection in humans each year in the
United States, with the highest reported incidence from California (4). Efforts to
reduce the number of infections have focused on vaccination of cattle herds. In 2008,
the USDA reported that all 50 states were Brucella Class Free, although sporadic cases
have altered the status of some states since then. Brucella continues to be present in
ferel swine, bison and the elk population of this country.
Infections are seen in essentially two patient populations. The first is individuals who
work with animals, which have not been vaccinated against brucellosis. This patient
population includes farmers, veterinarians, and slaughterhouse workers. B. abortus
(cattle) and B. suis (pigs) are the agents most likely to cause infections in this group of
3
LABORATORY GUIDELINES
FOR
SUSPECTED AGENTS OF
BIOTERRORISM
AND
EMERGING INFECTIOUS DISEASES
Brucella species
American Society for Microbiology (ASM)
Revised March 2016.
For latest revision, see web site below:
https://www.asm.org/Articles/Policy/Laboratory-Response-Network-LRN-Sentinel-Level-C
Mary K. York, Ph.D.
MKY Microbiology
Consultants Walnut Creek,
ASM Subject Matter Experts: CA
ASM Sentinel Laboratory
Peter H. Gilligan, Ph.D. Protocol Working Group
University of North Carolina Hospitals/
Clinical Microbiology and Immunology Labs
Chapel Hill, NC
,Vickie Baselski, Ph.D. Barbara Robinson-Dunn, Ph.D.
University of Tennessee at Department of Clinical
Memphis Memphis, Pathology
APHL Advisory Committee
TN Beaumont Health System
Royal Oak, MI
Patricia Blevins, MPH
BRobinson-
San Antonio Metro Health District
David Craft, Ph.D.
Laboratory
Penn State Milton S. Hershey
Medical Center Michael A. Saubolle, Ph.D.
Hershey, PA Banner Health System
Erin Bowles
Phoenix, AZ
Wisconsin State Laboratory of Hygiene
Peter H. Gilligan, Ph.D. m
University of North Carolina
Hospitals/ Christopher Chadwick, MS
Susan L. Shiflett
Association of Public Health Laboratories
Clinical Microbiology and Michigan Department of
Immunology Labs
Community Health
Chapel Hill, NC Lansing, MI
Mary DeMartino, BS,
MT(ASCP)SM
State Hygienic Laboratory at the University of
Larry Gray, Ph.D. Alice Weissfeld, Ph.D.
Iowa
TriHealth Laboratories and Microbiology Specialists Inc.
University of Cincinnati College Houston, TX
of Medicine Cincinnati, OH alice@microbiologyspecialists.c Harvey Holmes, PhD
om Centers for Disease Control and
Prevention
Major Todd Kijek, Ph.D. David Welch, Ph.D.
US Army Medical Research Medical Microbiology Kara MacKeil
Institute for Infectious Diseases Consulting Association of Public Health Laboratories
Ft. Detrick, MD Dallas, TX
Chris Mangal, MPH
Michael J. Loeffelholz, Ph.D. Mary K. York, Ph.D. Association of Public Health
Department of Pathology MKY Microbiology Consultants Laboratories
Univ. Texas Medical Branch Walnut Creek, CA
Galveston, TX
Amanda Moore, BS
South Carolina Department of
Coordinating Editor: Health and Environmental Control
Judith Lovchik, Ph.D.
Indiana State Department of
James W. Snyder, Ph.D.
Health Laboratories
University of Louisville
Indianapolis, IN James Rudrik, PhD,
Louisville, KY
Michigan Department of Community
Health
Scott W. Riddell, Ph.D.
Department of Pathology Administrative Support
SUNY Upstate Medical Maureen Sullivan, MPH Minnesota
University Kimberly E. Walker, Ph.D. Department of Health
Syracuse, NY American Society for
Microbiology
2
, PREANALYTICAL CONSIDERATIONS
I. PRINCIPLE
A. Introduction
Brucella is a fastidious, aerobic, small, gram-negative coccobacillus that is neither motile
nor spore-forming (11, 19). Brucella is considered an “Overlap select agent” because it
not only has the potential to pose a threat to public health and safety, but it also poses a
threat to animal health and animal products.
This procedure describes the steps to rule out, recognize, and presumptively identify
this organism in clinical specimens in Sentinel Clinical Laboratories. Such laboratories
are defined as those certified to perform high complexity testing under the Clinical
Laboratory Improvement Amendments of 1988 (CLIA) by the Centers for Medicare
and Medicaid Services for the applicable Microbiology specialty. Laboratory in-house
testing includes Gram stains, and at least one of the following: lower respiratory tract,
wound or blood cultures.
Sentinel Clinical Laboratories are not required to register with the Select Agent
Program to conduct diagnostic testing for Select Agents, both Tier I and non-Tier 1.
Testing for Select Agents may be performed by laboratories as long as the laboratory
follows the policies listed in the reporting section of this document when a Select
Agent cannot be ruled out. Consult with your designated LRN Reference Laboratory
or refer to the CDC Division of Select Agents and Toxins website at
http://www.selectagents.gov for questions.
NOTE: To identify Brucella to the genus level some biochemical testing is needed,
however, automated systems and manual multi-test kit identifications have no place in
the identification of the organism, due to its minimal reactivity, creation of aerosols in
performance of the testing, the danger of misidentification due to its close relation to
other organisms that are positive for only a few tests, and the easy, rapid method to
presumptively identify the organism without use of system identifications. Clinically,
rapid identification to the genus level is adequate to initiate therapy, and the type of
Brucella species involved does not alter the therapy.
B. Geographic distribution
There are between 80 and 120 cases of Brucella infection in humans each year in the
United States, with the highest reported incidence from California (4). Efforts to
reduce the number of infections have focused on vaccination of cattle herds. In 2008,
the USDA reported that all 50 states were Brucella Class Free, although sporadic cases
have altered the status of some states since then. Brucella continues to be present in
ferel swine, bison and the elk population of this country.
Infections are seen in essentially two patient populations. The first is individuals who
work with animals, which have not been vaccinated against brucellosis. This patient
population includes farmers, veterinarians, and slaughterhouse workers. B. abortus
(cattle) and B. suis (pigs) are the agents most likely to cause infections in this group of
3
