Smyser C F, Snoeyenbos G H
Avian Dis. 1976 Jul-Sep;20(3):545-51.
Direct culture and fluorescent antibody (FA) procedures were used for examining approximately 6200 samples of poultry floor or nest litter for salmonellae. Each sample was cultured directly in tetrathionate brilliant-green (TBG) broth, incubated at 42 C for 48 hours, and plated on brilliant green (BG) agar. The FA procedure was conducted from pools of 4-8 samples prepared by transferring 1 ml supernatant from TBG at 24 hr into 50 ml of either gram-negative (GN) broth, selenite cystine, or selenite brilliant-green sulfapyridine. After 6 hr of incubation at 42 C, BG agar plates were streaked from each pool, and smears were prepared for FA. Salmonella was isolated by plating from 268 of 854 pooled units. Recovery was 97% by direct culture, 86% from GN broth, and 57% from the selenite secondary enrichments. In the FA procedure, GN broth was superior to either of the selenites as the secondary medium from which to prepare smears, giving more FA-positive pools, a higher percentage of confirmed positives, and a lower percentage of false negative pools. FA examination of litter samples can be expedited by using sample pools. A major deficiency of the FA technique was the high percentage of false-positive reactions as judged by inability to isolate salmonellae. From litter cultured directly in TBG, salmonellae were isolated from 97% of the total number of pools culturally positive by combined culture methods.
采用直接培养法和荧光抗体(FA)法检测约6200份家禽地面或窝巢垫料样本中的沙门氏菌。每个样本直接接种于四硫磺酸盐煌绿(TBG)肉汤中,于42℃培养48小时,然后接种于煌绿(BG)琼脂平板上。FA法是将24小时培养的TBG肉汤中的1ml上清液转移至50ml革兰氏阴性(GN)肉汤、亚硒酸盐胱氨酸肉汤或亚硒酸盐煌绿磺胺吡啶肉汤中,制备4 - 8个样本的混合样本。在42℃培养6小时后,从每个混合样本中划线接种于BG琼脂平板,并制备涂片用于FA检测。从854个混合样本单元中的268个样本中通过平板接种分离出沙门氏菌。直接培养法的回收率为97%,GN肉汤培养法为86%,亚硒酸盐二次增菌培养法为57%。在FA法中,作为制备涂片的二次培养基,GN肉汤优于任何一种亚硒酸盐培养基,可产生更多FA阳性混合样本、更高比例的确诊阳性样本和更低比例的假阴性混合样本。使用样本混合法可加快垫料样本的FA检测。FA技术的一个主要缺陷是,根据无法分离出沙门氏菌判断,假阳性反应的比例较高。通过联合培养法,从直接接种于TBG的垫料培养物中,在97%的培养阳性混合样本总数中分离出了沙门氏菌。