Investigations

1st investigations to order

FBC

Test
Result
Test

Iron deficiency anaemia, autoimmune haemolytic anaemia, and idiopathic thrombocytopenic purpura can be seen in common variable immunodeficiency (CVID). Lymphopenia is seen in CVID, severe combined immunodeficiency, intestinal lymphangiectasia, and haematological malignancy. Lymphocytosis is seen in haematological malignancy.

It is important to note that adult and infant reference ranges for total lymphocyte count differ.

Result

normal or reduced Hb; normal, reduced, or increased lymphocytes; normal or reduced platelets

serum IgG

Test
Result
Test

May be reduced in common variable immunodeficiency, severe combined immunodeficiency, transient hypogammaglobulinaemia of infancy, and hyper-IgM syndrome. Is normal in selective IgA deficiency. In IgG subclass deficiency, laboratory testing shows ≥1 of: reduced IgG1, reduced IgG2, reduced IgG3, or reduced IgG4 (total IgG may be normal).

First-line test for suspected immunodeficiency. Test is repeated to confirm result. Age-related reference ranges are used. Immunoglobulins in infants <1 year of age may be of maternal origin and therefore results will not reflect ability to synthesise immunoglobulins.

Result

normal or reduced

serum IgA

Test
Result
Test

May be reduced in common variable immunodeficiency, severe combined immunodeficiency, transient hypogammaglobulinaemia of infancy, hyper-IgM syndrome, and IgG subclass deficiency. May also be normal in IgG subclass deficiency. Is absent in selective IgA deficiency.

First-line test for suspected immunodeficiency. Test is repeated to confirm result. Age-related reference ranges are used. Immunoglobulins in infants <1 year of age may be of maternal origin and therefore results will not reflect ability to synthesise immunoglobulins.

Result

normal, reduced, or absent

serum IgM

Test
Result
Test

May be reduced in common variable immunodeficiency and severe combined immunodeficiency. May be normal in selective IgA deficiency and IgG subclass deficiency. Is increased in hyper-IgM syndrome.

First-line test for suspected immunodeficiency. Test is repeated to confirm result.

Result

normal, reduced, or increased

serum and urine electrophoresis

Test
Result
Test

Performed in older patients. Presence of paraprotein may indicate haematological malignancy and secondary hypogammaglobulinaemia.

Result

normal or presence of paraprotein

serum free light chains

Test
Result
Test

Detects excessive free light chains and changes in kappa:lambda ratio.

Complementary to serum and urine electrophoresis.

Result

normal or excessive free light chains

urine dipstick

Test
Result
Test

May be positive in nephrotic syndrome and infection.

Result

normal or presence of protein, leukocytes, nitrites

serum creatinine

Test
Result
Test

increased in some renal conditions resulting in nephrotic syndrome.

Result

normal or increased creatinine

serum albumin

Test
Result
Test

May be reduced in protein-losing enteropathy, malabsorption, nephrotic syndrome, and sepsis.

Result

normal or reduced

LFTs

Test
Result
Test

Alkaline phosphatase/gamma-glutamyl transferase (gamma-GT) may be increased in granulomatous common variable immunodeficiency disease.

Result

normal or increased alkaline phosphatase/gamma-GT

sputum culture

Test
Result
Test

If positive, it is typically for encapsulated bacteria (e.g., Streptococcus pneumoniae, Haemophilus influenzae), but it can also be positive for non-encapsulated Haemophilus.

Cryptosporidium may be detected in hyper-IgM syndrome.

Result

normal, or positive for encapsulated/non-encapsulated bacteria

stool culture

Test
Result
Test

Cryptosporidium may be detected in hyper-IgM syndrome.

Result

normal, or positive for Campylobacter/Giardia/Cryptosporidium

chest x-ray

Test
Result
Test

Useful for acute infection, bronchiectasis, lymphadenopathy, and granulomatous disease. Absent/small thymus or structural thymic abnormalities may be seen in severe combined immunodeficiency. Thymoma is seen in Good syndrome.[30] Pneumatocoeles may occur in hyper-IgE syndrome.

Less sensitive than CT at picking up mild degrees of bronchiectasis, but much less radiation exposure.[31]

Result

normal; or evidence of infection, lymphadenopathy, thymic mass, or chronic damage

Investigations to consider

flow cytometry: lymphocyte subsets

Test
Result
Test

Usually performed by referral centres. Offers information about the immune cell counts and percentages (B-, T-, and natural killer [NK] cells).[32] Absolute counts are the important measure. Age-related reference ranges must be used, as normal ranges can vary widely (e.g., between infants and adults).

X-linked agammaglobulinaemia: significantly reduced or absent B cells (cell surface markers CD19, CD20).

Severe combined immunodeficiency: significantly reduced or absent T cells; B and NK cells may be normal or reduced.

Autoimmune lymphoproliferative syndrome: elevated double CD4 and CD8 negative population.

Chronic lymphocytic leukaemia, lymphoma: monoclonal population.

Result

variable

specific antimicrobial antibodies

Test
Result
Test

Usually performed by referral centres. Looks for specific antimicrobial antibodies to common bacterial and/or viral antigens (e.g., antibodies to Streptococcus pneumoniae, tetanus, Haemophilus influenzae).

Result

specific antibody deficiency/primary disease: reduced or absent

test immunisation response

Test
Result
Test

Test immunisations are usually performed by referral centres when specific antibody levels are reduced. Repeat testing is done at 4 to 6 weeks.

It is essential to remember that live vaccines (e.g., bacillus Calmette-Guérin [BCG], measles-mumps-rubella [MMR], yellow fever) are avoided in patients with suspected or known severe immunodeficiency. This must be considered following a case-by-case risk/benefit analysis.

Result

normal or reduced

IgG subclasses

Test
Result
Test

Usually performed by referral centres. A less dynamic assessment of antibody response than specific antibody tests. Disadvantages include presence of several technical issues; poorly defined normal ranges in young children; and lack of sufficient data on development in early childhood.

A low IgG subclass may not be associated with clinically significant disease.

Result

normal or reduced

flow cytometry: protein expression

Test
Result
Test

Usually performed by referral centres when testing for Wiskott-Aldrich syndrome protein, Bruton tyrosine kinase protein expression in X-linked agammaglobulinaemia, and CD40 ligand expression in X-linked immunodeficiency with hyper-IgM.

Result

Normal, reduced, or absent

genetic testing

Test
Result
Test

Usually performed by referral centres to confirm underlying genetic mutation. Important for diagnosis, family screening, and antenatal counselling/diagnosis (e.g., for hyper-IgM syndrome, X-linked agammaglobulinaemia).

Demonstrating bi-allelic mutations in the ADA gene confirms severe combined immune deficiency caused by adenosine deaminase deficiency.

Genetic testing is likely to become clinically important with the rapidly increasing identification of genetic primary immunodeficiency disorders.

Result

normal or mutation

enzyme assays

Test
Result
Test

Usually performed by referral centres in cases of suspected severe combined immunodeficiency (SCID).

The adenosine deaminase level may be low or absent in SCID.[27]

Result

variable

abdominal ultrasound

Test
Result
Test

Performed to check for the presence of a spleen and rule out differential diagnosis of hyposplenism.

Look for hepatosplenomegaly: this may occur in haematological malignancy causing hypogammaglobulinaemia, and also in common variable immunodeficiency (reactive, granulomatous, infection, lymphoma).

Result

normal, splenomegaly/hepatomegaly, or hyposplenism

CT chest/sinuses

Test
Result
Test

Usually performed by referral centres. Useful to detect bronchiectasis, lymphadenopathy, and granulomatous disease.[31] Disadvantages include a much higher radiation dose than chest x-ray; some patients with hypogammaglobulinaemia may be radiosensitive (e.g., ataxia-telangiectasia, common variable immunodeficiency).

Result

normal; or evidence of infection, lymphadenopathy, or chronic damage

CT abdomen

Test
Result
Test

Usually performed by referral centres.

Result

normal; hepatosplenomegaly; lymphadenopathy

pulmonary function test

Test
Result
Test

Usually performed by referral centres. Obstructive defect typically seen in bronchiectasis. Low diffusion capacity (transfer factor) of the lung typically seen in granulomatous lung disease (part of common variable immunodeficiency).

Result

normal; obstructive defect; or reduced diffusion capacity

lymph node biopsy

Test
Result
Test

Lymphadenopathy may be infectious or lymphoproliferative.

Infection may occur in any hypogammaglobulinaemia.

Lymphoid hyperplasia, granulomatous disease, and lymphoma associated with common variable immunodeficiency.

Result

normal, granulomatous disease, lymphoid hyperplasia, infection, or lymphoma

small- and large-bowel biopsy

Test
Result
Test

Usually performed by referral centres.

Result

normal, coeliac-like enteropathy, or inflammatory bowel disease lesions; common variable immunodeficiency: nodular lymphoid hyperplasia

polymerase chain reaction (PCR) for viral infection

Test
Result
Test

Usually performed by referral centres, as serology provides unreliable evidence of viral infection for hypogammaglobulinaemia. PCR provides direct demonstration of the infectious agent (e.g., hepatitis C).

Result

normal or positive

hepatitis B surface antigen, hepatitis B and C PCR

Test
Result
Test

Usually performed by referral centres before starting immunoglobulin replacement therapy, as there is a theoretical risk of infection. It is also performed as part of replacement immunoglobulin monitoring. Standard viral serology based on antibody detection is not useful in hypogammaglobulinaemia.

Result

negative

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