Two mRNA extraction methods were compared in this study to clarify the discrepancies found between authors regarding the presence of mRNA in inactivated Cryptosporidium parvum oocysts. Cryptosporidium parvum heat shock protein 70 (hsp70) mRNA extraction was performed by using oligo(dT)20-labeled magnetic beads or by incubating oocyst lysates with DNase I. Significant differences in mRNA recovery rates between these 2 techniques were observed when working on inactivated oocysts. We consistently detected hsp70 mRNA in oocysts heated at 60 C for 30 min and oocysts incubated in 10% formalin for 2 hr when using DNase I in the mRNA extraction procedure. In contrast, no mRNA was detected in such oocysts when magnetic beads were used for the mRNA extraction. The selective capture of long poly-A tail mRNA, when using oligo(dT)20-labeled magnetic beads, is proposed in this paper for explaining the discrepancies observed between the two mRNA extraction methods compared in this study. DNA decay in inactivated and aging oocysts makes quantitative polymerase chain reaction a potential alternative technique for assessing C. parvum oocyst viability status in environmental samples.
Skip Nav Destination
Article navigation
February 2001
RESEARCH NOTES|
February 01 2001
Relevance of Cryptosporidium parvum hsp70 mRNA Amplification as a Tool to Discriminate Between Viable and Dead Oocysts
J Parasitol (2001) 87 (1): 226–229.
Citation
P. Gobet, S. Toze; Relevance of Cryptosporidium parvum hsp70 mRNA Amplification as a Tool to Discriminate Between Viable and Dead Oocysts. J Parasitol 1 February 2001; 87 (1): 226–229. doi: https://doi.org/10.1645/0022-3395(2001)087[0226:ROCPHM]2.0.CO;2
Download citation file:
Sign in
Don't already have an account? Register
Client Account
You could not be signed in. Please check your email address / username and password and try again.
Could not validate captcha. Please try again.
Sign in via your Institution
Sign in via your InstitutionCiting articles via
SEROPREVALENCE OF TOXOPLASMA GONDII AND ASSOCIATED RISK FACTORS IN ONE-HUMPED CAMELS (CAMELUS DROMEDARIUS) IN QATAR
Sonia Boughattas, Hashim Alhussain, Ahmed Gawish, Nahla O. Eltai
SCOLICIDAL POTENCY OF BILE AGAINST ECHINOCOCCUS GRANULOSUS
İsa Caner Aydın, Noyan İlhan, Ahmet Şeker, Cem Batuhan Ofluoğlu, Fırat Mülküt, Nuri Emrah Göret, Kenan Çetin, Erdal Polat, Gökhan Aygün, Hasan Fehmi Kücük
SYNONYMIZATION OF PLACOBDELLA PICTA (VERRILL, 1872) (HIRUDINEA: GLOSSIPHONIIDAE) WITH DESCRIPTIONS OF TWO NEW SPECIES REVEALED BY MOLECULAR SPECIES DELIMITATION
Anna J. Phillips, Dennis J. Richardson, Charlotte I. Hammond, Brett C. Gonzalez, Eric A. Lazo-Wasem, William E. Moser
A RE-EVALUATION OF ZYGOCOTYLE (DIGENEA, PARAMPHISTOMOIDEA) BASED ON NEW GENETIC DATA SUPPORTS ITS SYNONYMIZATION WITH WARDIUS
Tyler J. Achatz, Dawn W. Cleveland, Sarah A. Orlofske, Robert C. Jadin, Jason Block, Lisa K. Belden, Hudson A. Pinto, Vasyl V. Tkach
DESCRIPTION OF A NEW SPECIES OF GYRINICOLA YAMAGUTI, 1938, FROM THE MONTEZUMA FROG, RANA MONTEZUMAE, IN THE TRANSITION ZONE BETWEEN THE NEARCTIC AND NEOTROPICS
Matthew A. Walker, Rogelio Aguilar-Aguilar, Virginia León-Règagnon, F. Agustín Jiménez