Major Histocompatibility Complex

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We have examined Ampdirect*TM (Shimadzu Corp., Japan) which is virtually PCR reaction buffer that eliminates the DNA extraction step and allows direct amplification from whole blood. In this study, we applied Ampdirect to the PCR-SSP method using the Dynal SSP kit (Dynal, Norway) to develop the most rapid DNA-hated HLA typing protocol by eliminating the time consuming step of DNA extraction. We tested this method for 5 healthy control donors and the primer specific PCR bands were detected as clearly as those obtained using purified DNA extracted by the conventional method, and thus identified alleles were consistent with those previously determined hy the PCR-RFLP method. The advantages of Ampdirect combined with the PCR-SSP method are not only to develop faster DNA typing procedure but also to reduce the chance to mix up samples by careless mistake, and further to eliminate technical difficulties associated with DNA purification steps. Ampdirect, hence, can he used as "PCR buffer" for amplification using whole blood, not purified DNA and it might be the most rapid and convenient technique for HLA DNA typing.

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