Fluorescence Quantitative Rt-pcr Method To Measure The Ebola Virus

Ebola hemorrhagic fever (EHF) is an acute hemorrhagic disease that is caused byEbola virus (EBOV), with a mortality rate as high as90%. EHF is one of the mostsevere infectious diseases to human, and poses a potential threat to public health. So far,there aren’t drugs of effective treatment and vaccines, and this disease isn’t found in ourcountry. So establishment of a method which is rapid, accurate and sensitive is veryimportant.In order to establish a rapid and accurate method for detection of EBOV, the primersand MGB probe used in MGB real-time RT-PCR were designed according to the EBOVNP gene sequence published in GenBank. One step MGB real-time RT-PCR and SYBRGreen Ⅰ real-time RT-PCR that detects Zaire and Sudan subtypes of EBOV wasestablished and optimized. The EBOV RNA that was transcribed in vitro was used astemplate. The sensitivity of the first method was found to reach1.0×103copies/μl andthe detection range was103-1010. The sensitivity of the second method was found toreach1.0×102copies/μl and the detection range was102-1010. This method did not reactwith RNA samples from Marburg virus (MARV), Dengue virus (DENV), Xinjianghemorrhagic fever virus (XHFV), Japanese encephalitis virus (JEV), Influenza virus(H1N1and H3N2) and Porcine reproductive and respiratory syndrome virus E genomicRNA. It has some innovation that the study quantitatively detected EBOV withreal-time RT-PCR and established One step MGB real-time RT-PCR for detection ofEbola virus Zaire and Sudan subtypes. The method would be useful for detection andinspection of EBOV in China.Susceptible animals of different species of Different areas are detected with twoestablished real-time RT-PCR methods. The results show that EBOV is found in ourcountry yet.