Comparison of concentration methods for detection of hepatitis A virus in water samples
- 87 Downloads
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 101 copies/μL and its concentration efficiency reached 79.45 ± 9.88%.
Keywordsconcentration filtration membrane hepatitis A virus mixed cellulose ester (MCE) membrane water samples
Unable to display preview. Download preview PDF.
- Beji-Hamza A, Khelifi-Gharbi H, Hassine-Zaafrane M, Della Libera S, Iaconelli M, Muscillo M, Petricca S, Ciccaglione AR, Bruni R, Taffon S, Equestre M, Aouni M, La Rosa G. 2014. Qualitative and quantitative assessment of hepatitis A virus in wastewaters in Tunisia. Food Environ Virol, 6: 246–252.CrossRefPubMedGoogle Scholar
- Collier MG, Khudyakov YE, Selvage D, Adams-Cameron M, Epson E, Cronquist A, Jervis RH, Lamba K, Kimura AC, Sowadsky R, Hassan R, Park SY, Garza E, Elliott AJ, Rotstein DS, Beal J, Kuntz T, Lance SE, Dreisch R, Wise ME, Nelson NP, Suryaprasad A, Drobeniuc J, Holmberg SD, Xu F, Hepatitis AOIT. 2014. Outbreak of hepatitis A in the USA associated with frozen pomegranate arils imported from Turkey: an epidemiological case study. Lancet Infect Dis, 14: 976–981.CrossRefPubMedGoogle Scholar
- Gazette of the Ministry of Health of People’s Republic of China. 2012). http://govinfo.nlc.gov.cn/gtfz/zfgb/wsb/688748a/201204/ P020120416401125621405.pdf?classid=373). (In Chinese)Google Scholar
- Jiang W, Zhuang L, Li S, Zhou J, Tang G, Wang D. 2013. Gene type analysis of hepatitis A virus in four outbreak cases in Guizhou province, China, from 2008 to 2009. Mod Prev Med, 40: 4260–4264. (In Chinese)Google Scholar
- Schwab KJ, Deleon R, Sobsey MD. 1995. Concentration and purification of beef extract mock eluates from water samples for the detection of enteroviruses, hepatitis-A virus, and Norwalk virus by Reverse Transcription-PCR. Appl Environ Microb, 61: 531–537.Google Scholar
- Wang S. 2009. Master’s thesis. Huazhong University of Science and Technology, Wuhan, China. Study on the methods of membran adsorption-elution and immunomagnetic beads adsorption for concentrating virus in water.Google Scholar
- Zhang ML, Yang J, Zhao H, Zhu L, Zhao HB, Cai BL. 2009. Detection and quantification of hepatitis A virus in Tianjin coastal seawater of Bohai Bay. Environ Sci, 30: 1608–1613. (In Chinese)Google Scholar