Evaluation of high hydrostatic pressure effect on human adenovirus using molecular methods and cell culture.
Evaluation of high hydrostatic pressure effect on human adenovirus using molecular methods and cell culture.
复制标题
使用分子方法和细胞培养评价高静水压对人腺病毒的影响。
DOI：
10.1016/j.ijfoodmicro.2012.06.006
复制
发表时间：
2012
影响因子：
5.4
通讯作者：
D. Rodríguez
中科院分区：
文献类型：
--
作者：
K. Kovač;M. Bouwknegt;M. Diez;P. Raspor;M. Hernández;D. Rodríguez
人类腺病毒（hav）在人类粪便中传播，因此可污染环境水，并可能通过灌溉转移到食物中。因此，需要高效的水和食品灭活技术。高压水静压（High hydrostatic pressure， HHP）处理是一种非热、节能、快速的紧急灭活技术，在消除食品中的病原微生物方面得到了广泛的研究。我们将HHP应用于HAdV-2在水和细胞培养基（CCM）中，并通过感染性试验、实时PCR （qPCR）和事先用蛋白酶K和dna酶i进行酶处理的qPCR (ET-qPCR)，测量了对病毒传染性和基因组和衣壳完整性的影响。虽然较低的压力不能提供令人满意的失活水平，但估计400和600MPa处理在有效应用于93秒和45秒时，可将病毒传染性降低约6 log10个单位。分别（即不包括压力单元的上浮次数）。然而，即使施加更高的压力，病毒基因组仍保持完整。虽然酸性pH可以保护HAdV-2免受HHP的失活，但在CCM中添加1%蔗糖时，没有观察到气压保护作用。另一方面，在CCM中添加10mM cacl2，估计可以在较长的处理时间（10min）下保护HAdV-2免受HHP的侵袭。当病毒在瓶装矿泉水中处理时，与在CCM中相同处理相比，观察到明显更高的传染性降低。总之，HHP被证明能在5秒内有效地将HAdV-2的传染性降低至6.5 log10单位，从而有助于防止食物和水传播病毒的公共卫生保护。然而，它的精确效果依赖于基质，因此需要考虑特定于基质的评价，以确保在实践中可靠的失活。
Human adenoviruses (HAdV) are shed in human faeces and can consequently contaminate environmental waters and possibly be transferred to foods by irrigation. Therefore, efficient inactivation technologies for water and foods are needed. High hydrostatic pressure (HHP) processing is a non-thermal, energy-efficient and rapid emergent inactivation technology, which has been widely studied to eliminate pathogenic microorganisms in foods. We have applied HHP to HAdV-2 in water and cell culture medium (CCM) and measured the effect on virus infectivity and genome and capsid integrity, by using infectivity assay, real-time PCR (qPCR) and qPCR with prior enzymatic treatment (ET-qPCR) with Proteinase K and DNase I. While lower pressures did not provide satisfactory inactivation levels, 400 and 600MPa treatments were estimated to reduce virus infectivity by approximately 6 log10units when effectively applied for 93s and 4s, respectively (i.e., excluding come up times of the pressure unit). However, virus genome remained intact even when higher pressures were applied. While acidic pH protected HAdV-2 from inactivation with HHP, no baroprotective effect was observed when 1% sucrose was added to the CCM. On the other hand, 10mM CaCl2added to the CCM was estimated to protect HAdV-2 from HHP with longer treatment times (>10min). When virus was treated in bottled mineral water, significantly higher infectivity reduction was observed compared to the same treatment in CCM. In conclusion, HHP was shown to effectively reduce HAdV-2 infectivity up to 6.5 log10units within 4s and can thus contribute to public health protection for food- and water-borne virus transmission. However, its precise effect is matrix dependent and therefore matrix-specific evaluations need to be considered for assuring reliable inactivation in practice.