Department of Environmental Health and Institute of Health and Environment, School of Public Health, Seoul National University, Seoul, Korea.
J Food Prot. 2009 Dec;72(12):2547-52. doi: 10.4315/0362-028x-72.12.2547.
We investigated the possibility of using hollow-fiber ultrafiltration (HUF) for the simultaneous recovery of multiple microorganisms in food samples. MS2 bacteriophage, E. coli, Bacillus subtilis spores, and murine norovirus (MNV) were each inoculated into 5 liters of either distilled water (DW) or glycine elution buffer and then concentrated using hollow-fiber polysulfone ultrafilters. The resulting concentrates were further analyzed by either cultivation or TaqMan real-time reverse transcription PCR assay. The overall average recovery rates were 7.1% in DW and 17.1% in glycine elution buffer. When the virus, vegetative bacteria, and bacterial spores were simultaneously inoculated into DW, glycine, or Tris-HCl elution buffers, on average 16.8% of inoculated microorganisms were recovered by HUF. The addition of 3% beef extract blocking buffer to HUF increased the total recovery rate to 46.1%, with incremental recovery rates increasing sharply for B. subtilis spores and MNV. Use of HUF resulted in E. coli recovery rates of 68.0% on lettuce and 66.2% on ham and MNV recovery rates of 1.5% on lettuce and 5.8% on ham. Our study demonstrates that HUF can be effective at simultaneously recovering and concentrating diverse bacterial and viral pathogens from foods.
我们研究了使用中空纤维超滤（HUF）同时从食品样品中回收多种微生物的可能性。将 MS2 噬菌体、大肠杆菌、枯草芽孢杆菌孢子和鼠诺如病毒（MNV）分别接种到 5 升蒸馏水中（DW）或甘氨酸洗脱缓冲液中，然后使用中空纤维聚砜超滤器浓缩。通过培养或 TaqMan 实时逆转录 PCR 分析对所得浓缩物进行进一步分析。在 DW 和甘氨酸洗脱缓冲液中的总体平均回收率分别为 7.1%和 17.1%。当病毒、营养细菌和细菌孢子同时接种到 DW、甘氨酸或 Tris-HCl 洗脱缓冲液中时，HUF 平均回收了 16.8%的接种微生物。向 HUF 中添加 3%牛肉提取物阻断缓冲液可将总回收率提高到 46.1%，枯草芽孢杆菌孢子和 MNV 的回收率递增幅度明显。使用 HUF 可使大肠杆菌在生菜上的回收率达到 68.0%，在火腿上的回收率达到 66.2%，MNV 在生菜上的回收率达到 1.5%，在火腿上的回收率达到 5.8%。我们的研究表明，HUF 可以有效地从食品中同时回收和浓缩多种细菌和病毒病原体。
