en Diagnostic/screening methods and protocols Diagnostic/screening methods and protocols Original article Original article <strong>Introduction: </strong><em>Cryptococcus neoformans</em> is an encapsulated fungal pathogen that causes life-threatening meningoencephalitis in immunocompromised patients. This yeast secretes several potent immunogenic proteins by secretory vesicular mechanisms, such as HSP70 chaperone. <strong>Methods:</strong> The PCR-amplified <em>HSP70 </em>gene was cloned into a PET-28a(+) expression vector. The purified recombinant HSP70&nbsp; (rHSP70) was evaluated by western blotting using an anti-His Tag-HRP antibody and then used for immunization of a rabbit. The serum of the immunized rabbit was tested against the whole lysate of <em>C. neoformans</em> in ELISA. <strong>Results:</strong> The antibodies in the rabbit&rsquo;s serum recognized lysate of <em>C. neoformans</em> yeast. The highest antibody levels were achieved after the third booster injection. <strong>Conclusion:</strong> The rHSP70 showed to be a reliable candidate for the designing and development of an ELISA kit for early detection of cryptococcosis, and to screen a large number of specimens. Cryptococcus neoformans, Enzyme-Linked Immunosorbent Assay, Heat Shock Protein 70KD 60 64 http://jommid.pasteur.ac.ir/browse.php?a_code=A-10-234-2&slc_lang=en&sid=1 Shima Safaei shimasafaei@yahoo.com 0000-0002-2646-4132 No Department of Pathobiology, School of Paraveterinary Sciences, BU-Ali Sina University, Hamadan, Iran. Hossein Rezvan h.rezvan@basu.ac.ir 0000000293280245 Yes Department of Pathobiology, School of Paraveterinary Sciences, BU-Ali Sina University, Hamadan, Iran. Roohollah Fateh rfateh@gmail.com 0000-0002-2073-5552 No Department of Microbiology and Immunology, School of Medicine, Qom University of Medical Sciences, Qom, Iran. Mohammad Khalifeh Gholi mkhgh2000@gmail.com 0000-0003-0474-2477 No Department of Microbiology and Immunology, School of Medicine, Qom University of Medical Sciences, Qom, Iran.