DETECTION OF EHP (Enterocytozoon hepatopanaei) IN WHITELEG SHRIMP (Litopenaeus vannamei) BY POLYMERASE CHAIN REACTION (PCR) METHOD

• Main Author: FITRIAH FAISAL - NIM: 21115004, ANNISA
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/25662
• Institution: Institut Teknologi Bandung

<p align="justify">Since 2014, white feces disease (WFD) is one of the main emerging problems for whiteleg shrimp farming industries in Indonesia. This outbreak is known to be caused by Enterocytozoon hepatopenaei (EHP) infection to shrimp. EHP infection resulted in growth retardation to mass mortality in shrimp. To date, WFD can be detected by histology, in situ hybridization and PCR. This study aimed to produce specific and sensitive primers to detect the presence of EHP early in whiteleg shrimp by PCR. In this study, we isolated the DNA of EHP from infected whiteleg shrimp, then PCR targeting spore wall protein (SWP) from EHP as well as sensitivity and specifity testing. As a result, EHP can be isolated from infected shrimp. In this study, we designed 3 pairs of primers (SWP-EHP1, SWP-EHP2 and SWP-EHP3) targeting spore wall protein of EHP. The design of the primers were obtained from the alignment of the entire SWP EHP sequence contained in the NCBI database. Of the three set of primers designed, two could be used for EHP detection using PCR, with PCR products from primers SWP-EHP1 was 398 bp and from SWP-EHP3 primers was 415 bp, with an optimum annealing temperature of 480C. This research was followed up with cloning and sequencing to ensure that the PCR products were part of the EHP target. Primers sensitivity test results revealed that primers SWP-EHP1 could detect EHP to 7,74 x 102 copies while the primers SWP-EHP3 could detect up to 16,2 x 102 copies.<p align="justify">