Diagnosis of Brucella: Prepared By: Bahaa Mostafa Kamel 415

Diagnosis of Brucella
Prepared by:
Bahaa Mostafa Kamel 415
Diagnosis of Brucella
Symptoms and signs of brucellosis are unspecific; cultures and serology are usually
necessary for diagnosis. Some general laboratory findings might suggest the diagnosis
(leukopenia, relative lymphocytosis and pancytopenia). Slight elevation in liver enzymes is a
very common finding. The criterion standard test for diagnosis of brucellosis is the isolation
of the organism from the blood or tissues (bone marrow, liver aspiration).
 Laboratory Studies
 Organism isolation
The Brucella abortus organism can be isolated from fetal lymph nodes, placenta, milk,
vaginal mucus, uterine exudate or semen. Also any fluid can be cultured (synovial, pleural
and cerebrospinal). Recovery of the organism requires the use of enriched culture media
and incubation in 10% CO2.
Three sulphonated broth tubes are inoculated with the patient's blood
- A blood broth tube incubated aerobically.

- A blood broth tube incubated in air + 10% CO 2.
- A blood broth tube incubated anaerobically.
The sensitivity of blood cultures with improved techniques such as the Castaneda bottles is
further improved by the lysis-centrifugation technique. With these methods, the sensitivity
is approximately 60%.
Subcultures are still advised for at least 4 weeks; thus, if brucellosis is suspected, the
laboratory should be alerted to keep the cultures for 3-4 weeks, which is not done routinely
for most bacterial cultures.
Bone marrow culture is thought to be the criterion standard, since the reticuloendothelial
system holds a high concentration of brucellae.
CSF evaluation: This reveals a mild-to-modest lymphocytic pleocytosis in 88-98% of in

patients with neurobrucellosis. Protein levels are elevated in conjunction with normal
glucose levels.
 Serological tests
Serological testing is the most commonly used method of brucellosis diagnosis.

These tests detect antibodies present in serum, milk, whey, vaginal mucus and seminal
plasma. The serological diagnosis is considered unreliable when performed during the
period of 2 to 3 weeks before and after abortion or calving.
-Serum agglutination is considered the standard test at this time. The serum agglutination
test will detect non-specific antibodies as well as those that are specific for Brucella abortus
infection and vaccination.
-Prozone phenomenon (which is the inhibition of agglutination with low titers and the
presence of agglutination with high titers), may occur secondarily to hyperantigenemia and
formation of blocking antibodies (IgA) interfering with IgG or IgM, possibly leading to false-
negative results, so routine dilution of the serum beyond 1:320 would help to prevent such a
problem.
-Rose Bengal test (buffered plate antigen or card test), This is a rapid and simple screening
test that is useful for detecting early infections.
-Tray agglutination (TAT) and modified TAT are also popular. Titers of more than 1:160 in
conjunction with compatible clinical presentation is considered highly suggestive of
infection. Titers of more than 1:320 are considered to be more specific, especially in
endemic areas. Seroconversion and evolution of the titers can also be used for diagnosis.
-Complement Fixation Test, This test has good specificity and is the most definitive test at
this point in time aside for bacterial isolation. It is used to confirm agglutination test and in
cases where agglutination test is negative.
-ELISA (Enzyme Linked Immunosorbent Assay)
This test has been useful during eradication programs after vaccination has ceased and is
used for screening or as a supplemental test to the complement fixation test. The ELISA test
has superior sensitivity and reliably detects true negative results.
-Milk Ring Test (antibody detection in milk), this test is a

satisfactory and inexpensive test used for surveillance of dairy
herds for brucellosis.
-Polymerase chain reaction (PCR): PCR testing for brucellae is

a recent advance with promising potential. It would allow for
rapid and accurate diagnosis of brucellosis. Two major genetic
targets are the Brucella gene BCSP31 and the 16S-23S rRNA operon. The 16S-23S rRNA
operon has been shown in studies to be more reliable in terms of sensitivity but is not yet
widely used in clinical practice and needs more standardization .
Other promising tests include nested PCR, real-time PCR and PCR-ELISA, but the clinical role
for these tests remains to be defined.
-Brucellin test: it is an allergic skin test produced by intradermal injection of:
1. A culture filtrate of Brucella

2. Brucella extract (Brucellin)
3. Purified protein
The reaction resembles the tuberculin reaction, it gives rise to an edematous indurated area
of at least 5 mm after 48 hours.
 Imaging Studies
 Chest radiography
Radiographic findings are typically absent in brucellosis, even in patients with prominent
respiratory symptoms.
Findings observed in patients with active pulmonary involvement include hilar and
paratracheal lymphadenopathy, pulmonary nodules, pleural thickening, and pleural effusion.
 Spinal radiography
Radiographic findings in patients with osteoarticular disease occur later in the course of
illness, usually 2-3 weeks after the onset of symptoms.
In patients with sacroiliitis, the most commonly observed abnormalities include blurring of
articular margins and widening of the sacroiliac spaces.
Spondylitis-related abnormalities include anterosuperior vertebral angle epiphysitis, spinal

straightening, narrowing of the intervertebral disc spaces, end-plate sclerosis, and
osteophytes.
 Radionuclide scintigraphy
This study is more sensitive for revealing skeletal abnormalities, especially early in the
disease, when standard radiographic findings are usually normal.
Radionuclide scintigraphy may be especially helpful in distinguishing hip involvement from

sacroiliitis.
To facilitate prompt diagnosis, this study also may have a role in screening for newly onset
brucellosis and musculoskeletal symptoms.
 Histologic Findings
Histologic findings in brucellosis usually include mixed inflammatory infiltrates with

lymphocytic predominance and granulomas (in up to 55% of cases) with necrosis.
References
- Brucella - Molecular and Cellular Biology
- JOURNAL OF CLINICAL MICROBIOLOGY, Sept. 1997, Vol. 35-No. 9
- RESEARCH ARTICLE - INTERNATIONAL MICROBIOLOGY (2004) 7:53–58
- http://emedicine.medscape.com/article/213430-diagnosis
- Brucellosis in humans and animals - WHO/CDS/EPR/2006.7
Prepared By:
Bahaa Mostafa Kamel - No. 415

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Diagnosis of Brucella

- A blood broth tube incubated aerobically.

CSF evaluation: This reveals a mild-to-modest lymphocytic pleocytosis in 88-98% of in

Serological testing is the most commonly used method of brucellosis diagnosis.

-ELISA (Enzyme Linked Immunosorbent Assay)

-Milk Ring Test (antibody detection in milk), this test is a

-Polymerase chain reaction (PCR): PCR testing for brucellae is

1. A culture filtrate of Brucella

Spondylitis-related abnormalities include anterosuperior vertebral angle epiphysitis, spinal

Radionuclide scintigraphy may be especially helpful in distinguishing hip involvement from

Histologic findings in brucellosis usually include mixed inflammatory infiltrates with

- Brucella - Molecular and Cellular Biology

- JOURNAL OF CLINICAL MICROBIOLOGY, Sept. 1997, Vol. 35-No. 9

- RESEARCH ARTICLE - INTERNATIONAL MICROBIOLOGY (2004) 7:53–58

- Brucellosis in humans and animals - WHO/CDS/EPR/2006.7

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