Comparison of three guanidinine thiocyanate RNA extraction methods applied to shellfish extract for enteric virus detection

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Research Paper 01/01/2014
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Comparison of three guanidinine thiocyanate RNA extraction methods applied to shellfish extract for enteric virus detection

Karamoko Yahaya, Aitmhand Rajaa, Ennaji MM
J. Bio. Env. Sci.4( 1), 122-127, January 2014.
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Abstract

Human enteric viruses are transmitted through water and food. It’s therefore necessary to include the research of these agents in the food and water sanitary quality analysis. Most of these viruses are not culturable. Gene amplification methods such as NASBA and RT-PCR are used for routine analysis. These methods of detection are possible only if there is an adapted procedure for extracting nucleic acids especially RNA. We proposed here to evaluate the quality of the three RNA extraction protocols using the guanidine thiocyanate. Two of the considered extraction methods are based on the work of Chomczynski and Sacchi (method I and method II). A third one which is a modification in order to reduce the working time was also applied (method III). Extraction tests were performed on mussel extract after concentration with PEG / NaCl . The quality of extraction is evaluated by spectrophotometry on the criteria “yield ” and ” purity” . The results show that method I and method II are equivalent and suitable and the method III was not appropriate for this type of samples.

VIEWS 10

Arnal C, Ferre-Aubineau V, Besse B, Mignotte B, Schwartzbrod L, Billaudel S.1999. Comparison of seven RNA extraction methods on stool and shellfish samples prior to hepatitis A virus amplification. Journal of Virological Methods 77:17-26.

Atmar RL, Neill FH, Romalde JL, Le Guyader F, Woodley CM, Metcalf TG, Estes MK. 1995. Detection of Norwalk virus and hepatitis A virus in shellfish tissues with the PCR. Applied and Enviromental Microbiology 61,3014-3018.

Bianchi S, Vecchio AD, Vilariño ML, Romalde JL. 2011. Evaluation of different RNA-extraction kits for sensitive detection of hepatitis A virus in strawberry samples. Food microbiology 28, 38-42.

Boom R, Sor CJ, Salilmans MMM, Jansen CL, Wertheim-Van Dillen PME Van der Noordaa J.1990. Rapid and simple method for purification of nucleic acids. Journal of Clinical Microbiology 28,495–503.

Bosch A. 1998. Human enteric viruses in the water environment: a minireview. International Microbioliology 1, 191-196.

Bosch A, Sanchez G, Le Guyader F, Vanaclocha H, Haugarreau L Pinto RM. 2001. Human enteric viruses in Coquina clams associated with a large hepatitis A outbreak. Water Science and Technolology 43, 61-65.

Bouchriti N, Goyal S M.1993. Methods for the concentration and detection of human enteric viruses in shellfish: a review. New Microbiologica 16, 105-113.

Chomczynski P. 1993. A reagent for the single-step simultaneous isolation of RNA, DNA and proteins from cell and tissue samples. Biotechniques 15, 532-4.

Chomczynski P, Sacchi N. 1987. Analytical Biochemistry 162, 156-159.

Cook N. 2003. The use of NASBA for the detection of microbial pathogens in food and environmental samples. Journal of Microbiological Methods 53,165-174.

Gonzalez-Perez I, Armas Cayarga A, García de la Rosa I, Josefina González González Y. 2007. Homemade viral RNA isolation protocol using silica columns: A comparison of four protocols. Analytical biochemistry 360, 148-150.

Greene J. 1993. Isolation and purification of nucleic acids: In recombinant DNA methodology course, Bio-tract 2 Fall session 4, 1-4.

Jiménez-Clavero M A, Ley V, Gómez N, Sáiz JC. 2006. Detection of enteroviruses. In Food-Borne Pathogens p. 153-169. Humana Press.

Karamoko Y, Ibenyassine K, Ait Mhand R, Idaomar M, Ennaji MM.2005. Adenovirus detection in shellfish and urban sewage in Morocco (Casablanca region) by the polymerase chain reaction. Journal of Virological Methods 126, 135– 137

Kingsley DH, Meade GK, Richards GP .2002. Detection of both hepatitis A virus and Norwalk-like virus in imported clams associated with food-borne illness. Applied and Environmental Microbiology 68,3914-3918.

Le Guyader FS Atmar RL .2007. Viruses in shellfish. Perspectives in Medical Virology 17, 205-226.

Richards GP .1999. Limitations of molecular biological techniques for assessing the virological safety of foods. Journal of Food Protection 62,691-697.

Santos N, Gouvea V. 1994. Improved method for purification of viral RNA from fecal specimens for rotavirus detection. Journal of Virological Methods 46, 11-21.

de Silva Gesteira A, Micheli F, Ferreira CF, de Mattos Cascardo, JC. 2003. Isolation and purification of functional total RNA from different organs of cacao tree during its interaction with the pathogen Crinipellis perniciosa.Biotechniques 35, 494-501.

Weber K, Bolander ME, Sarkar G. 1998. PIG-B: a homemade monophasic cocktail for the extraction of RNA. Molecular Biotechnology 9, 73-77.