Pusterla Nicola, Mapes Samantha, Johnson Cara, Slovis Nathan, Page Allen, Gebhart Connie
Department of Medicine and Epidemiology, School of Veterinary Medicine, University of California, Davis, CA 95616, USA.
J Vet Diagn Invest. 2010 Sep;22(5):741-4. doi: 10.1177/104063871002200513.
The purpose of the current study was to compare the molecular detection rate of Lawsonia intracellularis between feces and rectal swabs collected from 42 foals with suspected equine proliferative enteropathy (EPE). Fecal samples and rectal swabs were processed for DNA purification by using an automated extraction system. The purified DNA was then analyzed by real-time polymerase chain reaction (PCR) for the presence of the aspartate ammonia lyase (aspA) gene of L. intracellularis. Absolute quantitation was calculated by using a standard curve for L. intracellularis and expressed as copy numbers of the aspA gene of L. intracellularis per microliter of purified DNA. The combined PCR detection rate for L. intracellularis was 90%, with 38 foals testing PCR positive in feces (33 samples), rectal swabs (32), or both (27). Six foals tested PCR positive only in feces, whereas 5 tested positive only in rectal swabs. Feces yielded a significantly higher aspA gene copy number of L. intracellularis than rectal swabs. Feces and rectal swabs tested PCR negative from 4 foals. In conclusion, the results showed that feces yielded similar numbers of PCR-positive results, with a higher L. intracellularis aspA gene load than rectal swabs. By analyzing dual samples, the PCR detection rate for L. intracellularis increased from 76% and 79% for rectal swabs and feces, respectively, to 90%. Rectal swabs should be considered as an alternative sample type for EPE-suspected patients with decreased or no fecal output.
本研究的目的是比较从42匹疑似马增生性肠炎(EPE)的马驹采集的粪便和直肠拭子中胞内劳森菌的分子检测率。粪便样本和直肠拭子使用自动提取系统进行DNA纯化处理。然后对纯化的DNA进行实时聚合酶链反应(PCR)分析,以检测胞内劳森菌天冬氨酸氨裂解酶(aspA)基因的存在。通过使用胞内劳森菌的标准曲线计算绝对定量,并表示为每微升纯化DNA中胞内劳森菌aspA基因的拷贝数。胞内劳森菌的联合PCR检测率为90%,38匹马驹在粪便(33份样本)、直肠拭子(32份)或两者(27份)中PCR检测呈阳性。6匹马驹仅粪便PCR检测呈阳性,而5匹马驹仅直肠拭子检测呈阳性。粪便中胞内劳森菌的aspA基因拷贝数显著高于直肠拭子。4匹马驹的粪便和直肠拭子PCR检测均为阴性。总之,结果表明,粪便产生的PCR阳性结果数量相似,胞内劳森菌aspA基因载量高于直肠拭子。通过分析双份样本,胞内劳森菌的PCR检测率从直肠拭子的76%和粪便的79%分别提高到90%。对于粪便排出减少或无粪便的疑似EPE患者,直肠拭子应被视为一种替代样本类型。