A single platform image cytometer for resource-poor settings to monitor disease progression in HIV infection.

BACKGROUND

For resource-poor countries, affordable methods are required for enumeration of CD4(+) T lymphocytes of HIV-positive patients. For infants, additional determination of CD4/CD8 ratio is needed.

METHODS

We determine the CD4(+) and CD8(+) T lymphocytes as the CD3(+)CD4(+) and CD3(+)CD8(+) population of blood cells. Target cells are CD3-immunomagnetically separated from the whole blood, and CD4-Phycoerythrin and CD8-PerCP immunofluorescently labeled. A point-of-care single platform image cytometer was developed to enumerate the target CD3(+)CD4(+) and CD3(+)CD8(+) populations. It has light-emitting diodes illumination, is fully computer-controlled, operates from a 12 V battery, and was designed to be cheap and easy-to-handle. Target cells are imaged on a CCD camera and enumerated by an image analysis algorithm. The cytometer outputs the absolute number of CD4(+) and CD8(+) T lymphocytes/microl and CD4/CD8 ratio.

RESULTS

The quality of the cell images obtained with the cytometer is sufficient for a reliable enumeration of target cells. The image cytometer achieves an accuracy of better than 10% in the range of 50-1700 cells/microl. Analysis of blood samples from HIV patients yields a good agreement with the TruCount method for CD4 and CD8 count and CD4/CD8 ratio.

CONCLUSIONS

The image cytometer is affordable (component costs $3,000), compact (25 x 25 x 20 cm(3)), and uses disposable test materials, making it a good candidate to monitor progression of immunodeficiency disease in resource-poor settings.