A simple filter paper-based method for transporting and storing Enterocytozoon hepatopenaei DNA from infected Litopenaeus vannamei tissues

J Invertebr Pathol. 2020 Jan:169:107305. doi: 10.1016/j.jip.2019.107305. Epub 2019 Dec 6.

Abstract

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.

Keywords: DNA; EHP infection; FTA matrix card; Flinter associate technology.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Aquaculture
  • DNA, Protozoan / analysis*
  • Enterocytozoon / genetics
  • Enterocytozoon / isolation & purification*
  • Penaeidae / parasitology*
  • Specimen Handling / instrumentation
  • Specimen Handling / methods*

Substances

  • DNA, Protozoan

Supplementary concepts

  • Enterocytozoon hepatopenaei