Preservation of Giardia cysts in stool samples for subsequent PCR analysis.
Genotyping of Giardia duodenalis cysts in faecal samples has become a regularly employed tool by researchers investigating different aspects of the epidemiology and pathology of Giardia infection in human and animal populations. However, such investigations are often limited to some extent by lack of PCR amplification from a proportion of the samples, and this often seems to be associated with the storage medium used for the samples. Various different storage media have been used in different studies, but investigation of which storage media are most appropriate and which may compromise subsequent PCR investigations has not been systematically explored to date. In this study, 4 different, commonly used storage media were investigated for their effects over time on subsequent PCR amplification of DNA from Giardia cysts in stool samples. Microscopic examination of the samples and real-time PCR were used to investigate 7 different samples over a period of 3 months. Our findings indicate that storage in ethanol or potassium dichromate at 4 degrees C gave the best results and, that if immunomagnetic separation was used prior to PCR (as may be appropriate for samples with low cyst numbers), then storage in potassium dichromate gave the best results.
Wilke
Hans
H
Laboratory of Medical Microbiology Twente Achterhoek, 7500 AJ Enschede, The Netherlands. hwilke@labmicta.nl
Robertson
Lucy J
LJ
D000818:Animals; D003560:Cysts / Q000235:genetics*; D005243:Feces / Q000469:parasitology*; D005404:Fixatives / Q000494:pharmacology; D005838:Genotype; D005872:Giardia / Q000235:genetics* / Q000302:isolation & purification*; D006801:Humans; D010274:Parasitology / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods; D013048:Specimen Handling / Q000379:methods*; D013696:Temperature; D013997:Time Factors
D005404:Fixatives