Holoprosencephaly (HPE) is a cephalic disorder in which the prosencephalon (the forebrain of the embryo) fails to develop into two hemispheres. Normally, the forebrain is formed and the face begins to develop in the fifth and sixth weeks of human pregnancy. PCR technique, culture and histopathological examination is used in the diagnosis of GTB in female infertility. PCR was found to be useful in diagnosing early disease as well as confirming diagnosis in clinically suspected cases.