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Background  The injection of rabies immune globulin (RIG) is of the utmost importance in the management of category III exposures to rabies-suspect animals. Because of the high cost and limited availability of existing RIG, one possible replacement for RIG is monoclonal antibodies (MAbs) against the rabies virus (RABV). Consequently, it is necessary to determine the neutralizing activity of the MAbs against rabies viruses, especially street rabies virus. However, the method to detect the neutralizing activity of MAbs against street rabies virus remains undefined.    Methods  To establish a method for detecting the neutralizing activity of MAbs against street rabies virus, we constructed a library consisting of 12 strains of street RABV from 11 provinces in China. Using this street RABV library and the Reed–Muench formula, we established a method for detecting the neutralizing titer of the MAbs. The reliability and repeatability of the method were evaluated by repeatedly measuring the neutralizing activity of a MAb and a post vaccination serum.    Results  A total of 12 strains of street RABV were chosen for inclusion in the street RABV library, which covered six Chinese lineages (China I–China VI) and grew to high titers in N2A cells (> 10 5  FFD 50 /ml). On the basis of the library, we constructed the method to detect the neutralizing activity of the MAbs. The results of repeatedly measuring the MAbs and positive serum showed excellent reliability and repeatability of the method established in this study.    Conclusions  This study established a street RABV library reflecting the epidemiological features of Chinese rabies viruses, which provides a platform for detecting the neutralizing activity of MAbs against rabies viruses circulating in China.    Electronic supplementary material  The online version of this article (10.1186/s40249-018-0500-x) contains supplementary material, which is available to authorized users.

Establishment of a Chinese street rabies virus library and its application for detecting neutralizing activity