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

• Published in: Journal of invertebrate pathology 2020-01, Vol.169, p.107305-107305, Article 107305
• Main Authors: Karthikeyan, K., Saranya, Ravi, Bharath, Raja, Vidya, R., Itami, Toshiaki, Sudhakaran, Raja
• Format: Article
• Language: English
• Subjects: Animals; Aquaculture; DNA; DNA, Protozoan - analysis; EHP infection; Enterocytozoon - genetics; Enterocytozoon - isolation & purification; Flinter associate technology; FTA matrix card; Penaeidae - parasitology; Specimen Handling - instrumentation; Specimen Handling - methods
• ISSN: 0022-2011; 1096-0805
• DOI: 10.1016/j.jip.2019.107305

We synthesize novel poly(aryl ether ketone)s bearing flexible alkyl segments and pendant anthracene groups through nucleophilic copolycondensation coupled with post-polymerization functionalization. After the photodimerization of pendant anthracene units, the resultant polymers exhibit enhanced shape memory performance with satisfying shape fixity ratio and shape recovery ratio, attributed to their photocrosslinked polymeric network architecture. [Display omitted] •FTA matrix card can store the DNA of Enterocytozoon hepatopenaei.•Recovery of microsporidian DNA was efficient up to 6-months post spotting.•FTA extracted DNA quality was superior to that of other methods.•The present method facilitates easy transportation and extraction of EHP DNA. 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.