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首页> 外文期刊>Applied Microbiology >Evaluation of the Porcine Gastric Mucin Binding Assay for High-Pressure-Inactivation Studies Using Murine Norovirus and Tulane Virus
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Evaluation of the Porcine Gastric Mucin Binding Assay for High-Pressure-Inactivation Studies Using Murine Norovirus and Tulane Virus

机译：使用鼠诺如病毒和杜兰病毒进行高压灭活研究的猪胃粘蛋白结合试验的评估

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We compared the results of high-hydrostatic-pressure (HHP) inactivation of murine norovirus type 1 (MNV-1) and Tulane virus (TV) obtained by a porcine gastric mucin binding assay followed by quantitative reverse transcription-PCR (referred to here as the PGM-MB/PCR assay) and a plaque assay and evaluated HHP inactivation of a human norovirus (HuNoV) genogroup I genotype 1 (GI.1) strain and a HuNoV GII.4 strain by using the PGM-MB/PCR assay. Viruses were treated at different pressure levels for 2 min at 4 or 21°C in culture medium of neutral pH and in culture medium of pH 4 at 21°C. The log reductions of infectious MNV-1 and TV particles caused by HHP were assessed using the PGM-MB/PCR and plaque assays, while the log reductions of HuNoVs were assessed by the PGM-MB/PCR assay only. For TV and MNV-1, the two pressure inactivation curves obtained using the plaque and PGM-MB/PCR assays were almost identical at ≤2-log-reduction levels regardless of the treatment temperature and pH. Further increasing the pressure over the 2-log-reduction level resulted in higher log reductions of TV and MNV-1, as assessed by the plaque assay, but did not increase the log reductions, as assessed by the PGM-MB/PCR assay. HHP treatments could achieve maximum reductions of ～3 and 3.5 log units for GI.1 and GII.4, respectively, as assessed by the PGM-MB/PCR assay. On the basis of these results, it can reasonably be concluded that the PGM-MB/PCR assay would very likely be able to estimate HHP inactivation of HuNoV at ≤2-log-reduction levels. It would also likely conservatively quantify HHP inactivation of the GI.1 strain at 2- to 3-log-reduction levels and the GII.4 strain at 2- to 3.5-log-reduction levels.
机译：我们比较了通过猪胃粘蛋白结合测定，然后通过定量逆转录PCR（获得的结果）获得的鼠1型诺如病毒（MNV-1）和杜兰病毒（TV）的高静水压（HHP）灭活的结果（PGM-MB / PCR分析）和噬菌斑分析，并通过PGM-MB / PCR分析评估了人类诺如病毒（HuNoV）基因组I基因型1（GI.1）和HuNoV GII.4菌株的HHP灭活。在中性pH的培养基中和在21°C的pH 4的培养基中于4或21°C以不同的压力水平处理病毒2分钟。使用PGM-MB / PCR和噬菌斑分析评估了由HHP引起的感染性MNV-1和TV颗粒的对数减少，而仅通过PGM-MB / PCR分析评估了HuNoV的对数减少。对于TV和MNV-1，无论处理温度和pH值如何，使用噬菌斑和PGM-MB / PCR分析获得的两条压力失活曲线在≤2-log降低水平下几乎相同。如通过噬菌斑测定所评估的，将压力进一步提高至超过2-log-减少水平导致TV和MNV-1的较高的对数减少，但是如通过PGM-MB / PCR测定所评估的，并未增加对数减少。通过PGM-MB / PCR测定，HHP处理对GI.1和GII.4的最大减少量分别为〜3和3.5 log个单位。根据这些结果，可以合理地得出以下结论：PGM-MB / PCR分析很有可能能够估计HuNoV在≤2-log降低水平下的HHP失活。它也可能保守地将GI.1菌株的HHP灭活定量降低2至3个对数，而将GII.4菌株的HHP灭活定量降低2至3.5个对数。

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