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Chicken anemia virus (CAV) infection has been reported in various poultry industries worldwide. Since CAV infection is becoming increasingly prevalent, especially in local chickens of China, rapid CAV detection has become essential. The conventional diagnostic methods are time consuming and need special expertise. Therefore, in this study, we developed a specific and sensitive loop-mediated isothermal amplification (LAMP) assay for CAV detection by using multiple sequence alignment of VP2. This assay was performed at 61°C for 1 h, and there was no non-specific reaction to common avian disease viruses. The detection limit was 65 copies of viral DNA; thus, this assay showed similar sensitivity to quantitative polymerase chain reaction (qPCR) but it was more sensitive than conventional PCR. Moreover, this assay was performed using clinical samples. The LAMP assay results were 83.6% correlated to the PCR results of the clinical samples, indicating that this method is an effective tool for the rapid detection of CAV. [ABSTRACT FROM AUTHOR]