Département Lagons, Ecosystèmes et Aquaculture Durable en Nouvelle-Calédonie, IFREMER, Station de St. Vincent, B.P. 2059, 98846, Nouméa cedex, Nouvelle-Calédonie, France.
Microb Ecol. 2012 Jan;63(1):127-38. doi: 10.1007/s00248-011-9951-3. Epub 2011 Oct 15.
Two recurring syndromes threaten the viability of the shrimp industry in New Caledonia, which represents the second largest export business. The "Syndrome 93" is a cold season disease due to Vibrio penaeicida affecting all shrimp farms, while the "Summer Syndrome" is a geographically restricted vibriosis caused by a virulent lineage of Vibrio nigripulchritudo. Microbiological procedures for diagnosis of these diseases are time-consuming and do not have the ability to discriminate the range of virulence potentials of V. nigripulchritudo. In this study, we developed a multiplex PCR method to simultaneously detect these two bacterial species and allow for pathotype discrimination. The detection limits of this assay, that includes an internal amplification control to eliminate any false-negative results, were determined at 10 pg purified DNA and 200 cfu/ml. After confirming the effectiveness of our method using experimentally infected animals, its accuracy was compared to standard biochemical methods during a field survey using 94 samples collected over 3 years from shrimp farms encountering mortality events. The multiplex PCR showed very high specificity for the detection of V. penaeicida and V. nigripulchritudo (inclusivity and exclusivity 100%) and allowed us to detect the spreading of highly pathogenic isolates of V. nigripulchritudo to a farm adjoining the "Summer Syndrome area." This assay represents a simple, rapid, and cost-effective diagnostic tool for implementing timely risk management decisions but also understanding the seasonal and geographical distribution of these pathogens.
两种反复发作的综合征威胁着新喀里多尼亚虾业的生存能力，虾业是该国第二大出口产业。“93 综合征”是一种冷季疾病，由创伤弧菌引起，影响所有虾场；“夏季综合征”则是一种地理局限的弧菌病，由毒性很强的创伤弧菌引发。诊断这些疾病的微生物程序既耗时又无法区分创伤弧菌的各种潜在毒力。在这项研究中，我们开发了一种多重 PCR 方法，可同时检测这两种细菌，并能区分它们的病原类型。该方法的检测限为 10pg 纯化 DNA 和 200cfu/ml，包含一个内部扩增对照以消除任何假阴性结果。在用实验感染动物确认方法的有效性后，我们在为期 3 年的时间里，对虾场发生死亡率事件的 94 个样本进行了现场调查，用该方法与标准生化方法进行了准确性比较。该多重 PCR 对创伤弧菌和创伤弧菌的检测具有很高的特异性（包容性和排他性均为 100%），并能检测到高致病性创伤弧菌分离株向“夏季综合征”区域附近的虾场传播。该检测方法是一种简单、快速且具有成本效益的诊断工具，可用于及时做出风险管理决策，也有助于了解这些病原体的季节性和地理分布。
