Infectious hypodermal and hematopoietic necrosis virus (IHHNV) is one of the most serious diseases of shrimp,and has a wide host range. IHHNV is the smallest of all known shrimp viruses and is a non-enveloped, linear, single-strandedDNA virus in Brevidensovirus Densovirinae Parvoviridae. This disease causes serious economic losses worldwide. IHHNV ishighly pathogenic and is associated with high Litopenaeus stylirostrismortality rates. IHHNV causes chronic infection, slowgrowth, lesions in the forehead sword, antenna, and the cephalothoracic and abdominal shell of Litopenaeus vannamei. SinceIHHNV was first identified, many studies have established diagnostic methods, and epidemiological surveys have been conducted,but the pathogenic mechanisms of IHHNV infection remain relatively unknown. In this study, we identified putativehost cell receptors for the IHHNV capsid protein (CP) in L. vannamei gill membranes. DNA was extracted fromIHHNV-infected L. vannamei (Sheyang isolate) and preserved at the Microbiology and Immunology, Preventive VeterinaryLaboratory of Nanjing Agricultural University. The IHHNV CP gene was amplified by polymerase chain reaction, and theIHHNV CP BL21-pET-28a-CP prokaryotic expression vector was prepared. The protein was expressed primarily in the precipitate.The CP in the precipitate was purified using a HisTrap™ HP column and then used to prepare a polyclonal antibody.The antibody titer reached 1︰51200 after three immunizations. A Western blot analysis demonstrated that the polyclonal CPantibody bound specifically to CP. Then, gill membrane proteins from uninfected L. vannamei were obtained from a homogenateand ultracentrifuged to obtain a uniform gill membrane protein distribution. A virus overlay protein binding assay(VOPBA) and a His pull-downassaywere carried out to localize the IHHNV CP receptors. VOPBA is a classic method used toidentify virus receptors and has been used to identify the host cell receptors for White Spot Syndrome Virus and Yellow HeadVirus. The suspected protein strips were analyzed by mass spectrometry. Results of the two assays indicated that signal transducerand activator of transcription (STAT), heat shock protein 90 (HSP90), prophenoloxidase-2, and the Na+/K+-ATPase alphasubunit participate in IHHNV infection. These four proteins have various biological functions, such as interferon signaltransduction, protection against stress, defense, and maintenance of osmotic balance. These proteins may interact with theIHHNV CP and are associated with penetration and the cytopathic effects caused by IHHNV. Few studies have evaluated putativeIHHNV CP receptors, probably because of the lack of a mature shrimp cell line. We used traditional VOPBA to localizethe CP receptors from L. vannamei gill membrane proteins. Taken together, STAT, HSP90, prophenoloxidase-2, and theNa+/K+-ATPase alpha subunit are involved in IHHNV infection. These results will lay the foundation to identify the IHHNVinvasive mechanism. The specific effects of the four proteins remain to be studied in detail.