LAMP offers an alternative DNA amplification method to the polymerase chain reaction for applications to the ante-mortem saliva and CSF testing. The originators of the technique suggest that it amplifies with high specificity, efficiency and without the need for thermal cycling [38] . Amplification is achieved through the specific binding of two inner and two outer primers to the target sequence. The inner primers initiate strand synthesis whilst the outer primers displace the inner primers, allowing them to self-anneal to the nascent strand. This creates hairpin structures that trigger further strand synthesis that in turn lead to concatenation of the target sequence [38] . Polymerisation and strand displacement are achieved using a single enzyme, Bst 1 DNA polymerase. The technique is rapid, generating large quantities of target sequence within minutes. For the amplification of RNA viruses, a reverse transcription step is undertaken prior to the LAMP reaction. Primer sets have been successfully developed to detect a range of pathogenic viruses including West Nile virus [39] , Japanese Encephalitis virus [40] , Foot and Mouth Disease virus [41] and Chikungunya virus [42] . To assess the applicability of LAMP to the detection of rabies virus we designed a primer set using PrimerExplorer V4 software (Eiken Chemical Company Ltd., Japan) that can detect the Challenge Virus Standard (CVS) fixed strain of rabies virus (Table 6 ). In addition to the standard set of four primers, two further loop-binding primers have been added to increase the rate of strand displacement and synthesis [43] . The reverse transcription and LAMP reactions were undertaken simultaneously (RT-LAMP) in a single tube at 65uC using a www.plosntds.org thermostable reverse transcriptase, hence avoiding the step process inherent in an RT-PCR. Target amplification was monitored by the incorporation of the double stranded DNA binding fluorophore picogreen (Figure 1a) or by separation on a 1% agarose gel (Figure 1b) . At a constant temperature of 65uC, CVS RNA could be detected within 30 minutes. Development of RT-LAMP assays for use in diagnosis and surveillance is challenged by the considerable sequence variation observed within the rabies virus genome [44] that can frustrate specific primer design. Preliminary attempts at this suggest that multiple combinations of primers (up to 12 different primers) can lead to sensitive, rapid amplification of RABV genomes from a wide range of geographical locations. The use of isothermal amplification has the benefit of reducing the technological requirements of thermal cycling used in RT-PCR. This in turn offers the opportunity, when linked with lateral flow devices, to develop surveillance protocols where testing can take place in the field or in less sophisticated laboratories.