[Development of an extraction and concentration method for the detection of hepatitis A virus in different samples].
OBJECTIVE
To develop an extraction and concentration method for the detection of hepatitis A virus (HAV) in shellfish, water, serum and saliva samples by nested RT-PCR.


METHODS
HAV were artificially inoculated into the above samples, calm sample was extracted using glycine buffer pH9.5, PEG precipitation; water sample was PEG precipitated directly; then all the samples including serum and saliva samples were extracted using Trizol regent, followed by nested RT-PCR detection using primers from HAV VP1-2A region.


RESULTS
The detection limit for HAV in cultured cell lysis was 0.1TCID50; in water, serum or salva sample was 1TCID50 respectively, in calm sample was 1-10 TCID50. HAV RNA was detected in water and sera samples collected from the HAV outbreak region, sequenced and analysis.


CONCLUSIONS
The method developed here is convenient, specific and capable of detecting low levels of HAV in different samples, would be useful for diagnostic laboratories in order to perform HAV analysis in cases of foodborne infections or for molecular epidemiology investigation of HAV outbreaks.
Meng
Qing-Ling
QL
Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China.
Guo
Zi-Jie
ZJ
Cao
Jing-Yuan
JY
Bi
Sheng-Li
SL
D000818:Animals; D005591:Chemical Fractionation / Q000379:methods*; D005618:Fresh Water / Q000821:virology; D005821:Genetic Techniques; D006506:Hepatitis A / Q000175:diagnosis* / Q000821:virology; D030041:Hepatitis A virus / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D012463:Saliva / Q000821:virology; D044967:Serum / Q000821:virology; D012758:Shellfish / Q000821:virology
D012367:RNA, Viral