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A simple filter paper-based method for transporting and storing Enterocytozoon hepatopenaei DNA from infected Litopenaeus vannamei tissues
Saranya R, Bharath R, , Itami T,
Published in Elsevier BV
PMID: 31816302
Volume: 169
The microsporidian parasite Enterocytozoon hepatopenaei (EHP) causes a significant negative impact in shrimp aquaculture. A diagnostic procedure for detecting EHP in shrimp was developed, but transportation of the infected shrimp samples from the farm / hatcheries to the laboratory is burdensome and preservation of the tissues is problematic. Here, we developed a simple method of transporting nucleic acid without preservatives using the Flinders Technology Associates filter card (FTA matrix card; Whatman). DNA can be stored and extracted without the need for centrifugation and hazardous chemicals. EHP infected shrimp homogenate was spotted on a FTA matrix card and stored at room temperature. Storage stability was confirmed by analysis at different time points and we efficiently recovered DNA up to 6 months post spotting. The recovery efficiency of FTA-DNA was compared with the existing DNA extraction methods DNeasy® Blood & Tissue kit method and Guanidine hydrochloride method. The efficiency of extraction and sensitivity of the DNA in the FTA card confirmed that recovery of EHP-DNA from the FTA matrix was superior to with other methods. © 2019 Elsevier Inc.
About the journal
JournalData powered by TypesetJournal of Invertebrate Pathology
PublisherData powered by TypesetElsevier BV
Open AccessNo