Sample suitability for the detection of minor white cell populations (microchimerism) by polymerase chain reaction.
BACKGROUND
There is increasing use of highly sensitive testing with polymerase chain reaction (PCR) to study white cell microchimerism after transfusion and transplantation. This study investigated possible artifactual sources of allogeneic sample contamination before PCR testing.


METHODS
Quantitative Y-chromosome PCR was used to study microchimerism among transfused patients with sickle cell disease (SCD) and thalassemia by using residual specimens from the clinical laboratory. High levels of circulating male white cells among transfused patients with SCD but not thalassemia led to concern over the artifactual origin of male cells. To investigate, paired specimens were collected from 26 female SCD patients: one specimen underwent processing only for PCR, while the other underwent testing in the clinical laboratory before PCR as a process control. All laboratory instruments were also assessed for their ability to impart male allogeneic cells to aliquots of female blood.


RESULTS
Thirty-three (31%) of 107 SCD samples, but 0 of 20 thalassemia samples, gave a high-level PCR signal. One of 26 paired samples that was not exposed to clinical laboratory equipment had low-level PCR positivity while 10 of the 26 became strongly positive after testing on a blood cell analyzer and a reticulocyte analyzer. Sixteen of 32 female samples became positive after reticulocyte analysis, while none became positive after blood cell analysis. Samples from thalassemia patients tested PCR-negative because reticulocyte counts had not been performed.


CONCLUSIONS
Allogeneic cell contamination is common with clinical laboratory equipment. These samples may not be suitable for microchimerism studies. In addition to method controls, process controls should be employed where appropriate.
Reed
W
W
Blood Centers of the Pacific, San Francisco, California, USA.
Lee
T H
TH
Vichinsky
E P
EP
Lubin
B H
BH
Busch
M P
MP
D000755:Anemia, Sickle Cell / Q000235:genetics; D001803:Blood Transfusion; D004247:DNA / Q000032:analysis; D004340:Drug Contamination; D017743:Equipment Reuse; D005189:False Positive Reactions; D005260:Female; D006403:Hematologic Tests / Q000295:instrumentation*; D006801:Humans; D007962:Leukocytes / Q000737:chemistry*; D016555:Matched-Pair Analysis; D016133:Polymerase Chain Reaction; D013048:Specimen Handling; D013789:Thalassemia / Q000235:genetics; D018183:Transplantation Chimera / Q000235:genetics; D014998:Y Chromosome / Q000235:genetics
D004247:DNA