Since 1997,highly pathogenic avian influenza (HPAI) H5N1 viruses have caused serious outbreaks in poultry and markets,In human,overall mortality in HPAI H5N1 infection exceeds 60％,but human to human transmission is limited and has been only reported within family members[7,8].There is much concern as to whether H5N1 can enhance its transmission among humans through genetic variation.Further,there is an urgent need to discover the potential mutations in viral proteins that are responsible for inter-human transmission.
Hepatitis A virus is a pathogen associated with water pollution.Contaminated drinking water can cause hepatitis A outbreaks,lead to economic losses,and even threaten human lives.It is difficult to detect low levels of hepatitis A virus in water,so the virus must be concentrated in order to quantify it accurately.Here,we present a simple,rapid,efficient technique for the concentration and detection of hepatitis A virus in water.Our data showed that adding phosphate-buffered saline to the water,pre-filtering the water,and adding Trizol reagent directly to the filtration membrane can significantly improve concentration efficiency.Of three types of filtration membranes studied(mixed cellulose ester membrane,polyvinylidene fluoride membrane,and nylon membrane),the concentration efficiency using mixed cellulose ester membrane with a 0.1-μm pore size was the highest,reaching 92.62 ± 5.17%.This method was used to concentrate hepatitis A virus in water samples from Donghu Lake.Using SYBR Green real-time reverse transcription polymerase chain reaction analysis,the detection sensitivity of this method reached 10~1 copies/μL and its concentration efficiency reached 79.45 ± 9.88%.
