Multiplex One-step RT-PCR for Detection and Serotyping of Foot and Mouth Disease Virus in Balochistan

Foot-and-mouth disease (FMD) is a highly infectious and contagious disease of cloven hoofed animals. The disease is endemic in Pakistan caused by Foot-and-mouth disease virus (FMDV) serotype O, A and Asia-1. In order to control FMD in Pakistan, it is essential to have an accurate knowledge of circulating virus strains in the country that will help to select more specific vaccines. Balochistan province (Largest Province area wise) has an important geographical location, having common borders with Afghanistan and Iran that are also FMD endemic countries. Data regarding FMDV strain circulating in this province is scanty. Balochistan being the largest province of the country and having important geographical location needs to be explored for the detection of different serotypes circulating along with their lineages and sub lineages in order to control the disease as well as to access the routes of animal movement in the region, which could be the source of spread of virus. Furthermore, restriction of cross border movement is the need of time, along with the establishment of laboratories having capability for early and accurate diagnosis of the disease. This study suggests that there should be extensive sampling from this province for detection and typing of the virus and if possible, isolation of the circulating strains. This will help to identify Hot Spots in the region for regular monitoring against this alarming threat for the region. The serotype and strain identification will help to improve the current vaccines being used in the region. Sensitive, specific and quick diagnostic tools are needed to control the disease in FMD endemic regions, especially in the areas with limited facilities to diagnose the disease. In this paper, the situation of FMD in Balochistan province of Pakistan has been taken into consideration where FMD is endemic. Balochistan Province has limited diagnostic facilities in terms of infrastructure, equipment, kits, chemicals and reagents to perform all the techniques recommended by OIE, World Organisation for Animal Health, to detect and type the FMD virus. Most of the commercially available assays are too expensive to use for diagnosis with limited resources. This paper describes the optimization of multiplex one-step RT-PCR using specially designed primers for this region. This will serve as a milestone in early detection and serotyping of FMDV in Balochistan.