Investigations

1st investigations to order

blood culture

Test
Result
Test

Considered the mainstay of diagnosis.[90] The sensitivity of blood cultures depends largely on previous antibiotic use, the phase of the disease, and the method of culturing.[91][92][93]

Traditional culture sensitivity is improved by using sensitive methods such as lysis-centrifugation or specialised culture media (e.g., Castañeda's medium) and by extending culture duration to 6 weeks. Modern culture systems are more sensitive and usually become positive within 1 week, but subcultures should be maintained for up to 3 weeks.[93][Figure caption and citation for the preceding image starts]: Small pearly-white colonies of Brucella melitensis after prolonged culture on blood agarFrom the collection of Dr Nicholas J. Beeching; used with permission [Citation ends].com.bmj.content.model.Caption@262b6360

Result

positive growth of Brucella species

serological tests

Test
Result
Test

Cultures are not always available or successful, so patients are usually investigated using at least one serological test, typically an agglutination test or enzyme-linked immunosorbent assay (ELISA). Serological tests are still based on the traditional (Wright) standard agglutination test (SAT) or tube agglutination test (TAT), modified in some laboratories to be performed in small ELISA plates as the microagglutination test (MAT).[94]

The techniques suffer from lack of standardisation and are notoriously affected by the prozone phenomenon, caused by blocking IgA antibodies, yielding false negative results at low dilutions of patient serum.[1]

ELISA tests usually have high sensitivity but variable specificity.[84][94][96]

Rose Bengal agglutination tests, developed for veterinary practice, are used in some countries to screen human sera but require subsequent confirmatory testing.[95][94]

There is considerable cross-reactivity between Brucella species, as well as with other gram-negative bacteria; therefore, species diagnosis by serology is not reliable.[98]

Rapid point-of-care serological assays have been developed but require further evaluation before introduction into routine practice.

Routine serological tests are not effective for diagnosing or monitoring infection with B canis or with RB51 (the B abortusstrain used in animal vaccines).[77]​ Using a combination of serological tests may overcome the current limitations of testing.[120]

Result

positive agglutination titre of ≥160, or a fourfold or greater rise in titre between samples taken 2 weeks apart

cerebrospinal fluid (CSF) analysis

Test
Result
Test

Lumbar puncture is indicated in patients with neurological signs and symptoms to exclude meningoencephalitis. Serological tests on CSF are difficult to interpret, but the standard agglutination test is usually positive.[1][108][109]

Result

lymphocyte predominance with mildly raised protein and reduced or borderline glucose levels

cerebrospinal fluid (CSF) culture

Test
Result
Test

Rarely positive, but is improved by use of automated culture systems.[109]

Result

may show positive growth of Brucella species

synovial fluid analysis

Test
Result
Test

Indicated in all patients with joint effusion.

Result

lymphocyte predominance with raised protein and reduced glucose levels in cases of arthritis

synovial fluid culture

Test
Result
Test

Indicated in all patients with joint effusion. Frequently positive.

Result

positive growth of Brucella species

FBC

Test
Result
Test

Thrombocytopenia and/or anaemia can occur in 30% to 75% of infected patients.[111][112]

Leukopenia or leukocytosis occurs in 21.7% and 7.2% of patients, respectively.[111]

Result

anaemia, thrombocytopenia; leukopenia or leukocytosis

liver function tests

Test
Result
Test

Mild disturbance of transaminases considered a common finding.[1][84]

Result

mild elevation of transaminases

serum electrolytes

Test
Result
Test

​Hyponatraemia, associated with SIADH, has been reported in patients with brucellosis.[113]​ May also reveal other electrolyte derangements.

Result

hyponatraemia

Investigations to consider

bone marrow culture

Test
Result
Test

Bone marrow cultures have a greater positive yield than blood cultures, as the organism is intracellular and localised in the bone marrow, and may be considered in difficult cases (e.g., negative blood cultures, negative serology, and brucellosis suspected).[91]

Result

positive growth of Brucella species

tissue biopsy

Test
Result
Test

Clinically affected organs and tissues can be biopsied, particularly lymph nodes, liver, and, occasionally, synovium. This is partly to obtain material for culture to exclude tuberculosis.

Result

non-caseating granulomata

plain film x-rays of affected joints

Test
Result
Test

These changes in the axial skeleton and peripheral joints occur late in the disease.[1]

Result

small erosions near affected joints, moderate sclerosis of bone adjacent to infected joints, and little joint destruction or loss of joint space

chest x-ray

Test
Result
Test

Findings typically normal. Reserved for patients with pulmonary signs and symptoms.[88]​​

Result

normal, or pleural effusion and consolidation

bone scan

Test
Result
Test

Sensitive and may reveal subclinical joint infection.[115]

This study may be of use early in the disease, when abnormalities are usually not visible on plain film radiographs, and should be considered for patients with musculoskeletal manifestations. Furthermore, bone scan may help distinguish hip involvement from sacroiliitis.

Result

variable

CT or MRI scan of spine

Test
Result
Test

Useful for delineating infection of the spine and paraspinal tissues.

Result

variable

CT or MRI scan of head

Test
Result
Test

Rare findings in cases of central nervous system infection.

Result

may show intracranial collections, calcification, or hydrocephalus

Emerging tests

polymerase chain reaction (PCR) for detection and diagnosis of Brucella species

Test
Result
Test

Usually more rapid than culture and has sensitivity reported to approach 100%, with a specificity of 98.3%.[102]

May be particularly useful in patients with relapse or re-infection and has been used in trials to monitor progression of treated patients and assess relapse.[103]

However, PCR methods are still not standardised, are susceptible to contamination, and have yielded contradictory results with prolonged positivity in some settings, so their full potential has yet to be realised.[104]

Result

positive

matrix-assisted laser desorption/ionisation time of flight (MALDI-TOF) mass spectrometry

Test
Result
Test

There is increasing evidence for the use of MALDI-TOF mass spectrometry for rapid analysis and identification of Brucella species in blood and pure cultures; however, this is still considered an emerging test.[105][106][107]

Result

positive

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