Phenotypic and Genotypic Characterization of von Willebrand Factor Gene of Saudi Healthy Individuals in Eastern Province of Saudi Arabia

Abstract

Background:Vibrio parahaemolyticus is a gram-negative, facultative, anaerobic,halophilic bacterium that is widely disseminated in coastal, estuarine, andmarine environments. It is a seafood-borne pathogen that, in humans, cancause gastroenteritis. There are many clinical symptoms, such as diarrhea,abdominal pain, fever, and vomiting. Not all strains of V. parahaemolyticusare pathogenic, except, those are expressed tdh or trh genes. The polymerasechain reaction is the most popular molecular technique used to detect V.parahaemolyticus virulence genes. Recently, many studies have used anewly established technique known as immunomagnetic beads (IMB) toconcentrate the bacterium. This will increase the possibility of detecting V.parahaemolyticus from environmental, food, and clinical samples. Thisstudy aims to use PCR and IMB to detect V. parahaemolyticus in coastalwater samples.Method and result:From February 2018 to May 2018, seawater samples were collected fromfive different areas along the Arabian Gulf Coast in Saudi Arabia, includingAziziya Beach (AZB), Corniche alkhobar (KBC), Corniche alkhobar front(KBF), Half-Moon Beach (HMF), and Dammam corniche (DMC). In total,192 seawater samples were collected with measured physical parameters12such as pH, temperature, and TDS (total dissolved solids). Samples weretested to detect the presence of V. parahaemolyticus. IMB was used withTCBS and CHROMagar to increase the isolates of V. parahaemolyticus.Also, a molecular study was done by using PCR to confirm the identity of V.parahaemolyticus to the species level and identify virulence gene markers.ERIC PCR was performed to detect the similarities between V.parahaemolyticus strains. In total, 58 isolates were tested positive for V.parahaemolyticus after the use of TCBS and CHROMagar media with IMB,while the total number of isolates without IMB was only 9. To confirm theidentity of the species level, PCR targeting the toxR gene was applied and 28strains were detected. The pathogenicity of isolated strains was tested byusing PCR targeting tdh and trh genes, 23 isolates are tdh+ and trh- genesnot detected.Conclusion:The seawater samples were tested with and without IMB. The finding of thisstudy confirms the application of IMB in increase the detection level of V.parahaemolyticus. This study concludes that V. parahaemolyticus tdh+ ispresent in the analyzed samples of seawater collected from coastal areasalong the eastern coast of Saudi Arabia.