Rapid and sensitive PCR detection ofVibrio penaeicida, the putative etiological agent of Syndrome 93 in New Caledonia

Diseases of Aquatic Organisms

Jean Christophe Avarre¹  Denis Saulnier¹  Vincent Vonau¹  Gilles Le Moullac¹  Peva Levy¹  Dominique Ansquer¹ 

关键词: Syndrome 93;    Vibrio penaeicida;    PCR diagnosis;    Shrimp disease;    Vibriosis;    Penaeus (Litopenaeus) stylirostris · Small subunit rDNA gene;   

DOI  :  10.3354/dao040109

学科分类：生物科学（综合）

来源: Inter-Research

【 摘 要 】

ABSTRACT: Experimental infections of Penaeus (Litopenaeus) stylirostris were performed with a Vibrio penaeicida strain (AM101) isolated in New Caledonia from Syndrome 93 diseased shrimp. Cumulative mortalities resulting fromintramuscular injection or immersion of shrimp in bacterial suspensions demonstrated high virulence for this bacterial strain and suggested that V. penaeicida could be the etiological agent of Syndrome 93. The median lethal dose (LD₅₀)for AM101 was 1.3 x 10⁴ CFU (colony forming units) ml^-1 by immersion and less than 5 CFU shrimp^-1 by intramuscular challenge, with mortality outbreaks at 48 and 22 h after challenge,respectively. A polymerase chain reaction (PCR) detection assay using a primer set designed from the 16S ribosomal RNA gene of V. penaeicida was developed. It gave an expected amplicon of approximately 310 bp in ethidium bromide-stained agarosegels. The specificity of these primers was assessed with different Vibrio species. Furthermore, DNA extracted by the Chelex^TM method could be used to detect fewer than 20 cultured Vibrio cells in seawater or shrimp hemolymph bythis assay. It appears to be a reliable screening method for detecting V. penaeicida in shrimp and from the aquatic environment.

【 授权许可】

Unknown