DDx – no gowth or poor growth

· (blood culture)

· Suboptimal sample

· Hemolyzed

· Clotted

· No T cells

· DiGeorge

· Few lymphocytes

· Try using the buffy coat layer

· Patient on IV drugs

· Try washing cells with medium before culturing

· Patient pregnant

· Try using buffy coat layer

· Critically ill or deceased patient

· Specimen stored too long

· Poor culture conditions:

· Too much blood in culture

· Medium inadequate

· pH too basic

· mitotic stimulant was not added

· insufficient gas exchange

· microbial contamination

· incubator temperature too low or too high

· medium, serum, or PHA expired

· Harvest conditions suboptimal

· Cells lost during centrifugation

· Centrifugation not done prior to aspiration of supernatant

· Hypotonic solution improperly made

· Check pH

· Fixative improperly made

· Fixative ratio reversed

· Plastic from bottle caps contaminating

·

References:

· Barch MJ, Knutsen T, Spurbeck JL. The AGT Cytogenetics Laboratory Manual. 3rd ed. Lippincott Williams & Wilkins; 1997.