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首页> 外文期刊>Mayo Clinic Proceedings >Detection of vaccinia virus, herpes simplex virus, varicella-zoster virus, and Bacillus anthracis DNA by LightCycler polymerase chain reaction after autoclaving: implications for biosafety of bioterrorism agents.
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Detection of vaccinia virus, herpes simplex virus, varicella-zoster virus, and Bacillus anthracis DNA by LightCycler polymerase chain reaction after autoclaving: implications for biosafety of bioterrorism agents.

机译：高压灭菌后，通过LightCycler聚合酶链反应检测牛痘病毒，单纯疱疹病毒，水痘带状疱疹病毒和炭疽芽胞杆菌DNA：对生物恐怖主义剂的生物安全性的影响。

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OBJECTIVE: To determine whether autoclaving suspensions of vaccinia virus, herpes simplex virus (HSV), varicella-zoster virus (VZV), and Bacillus anthracis inactivate infectivity of these agents but allow detection of target DNA by LightCycler polymerase chain reaction (PCR). MATERIAL AND METHODS: Swabs were inserted into tubes containing serial 10-fold dilutions (10(-1) to 10(-5); 500 microL; 6 samples per dilution) of vaccinia virus, HSV, VZV, or a single suspension of 10(8) colony-forming units of B anthracis (2 samples). One half of the samples were autoclaved, and the remainder were not. An aliquot of each not autoclaved sample served as a control for infectivity. RESULTS: Autoclaving swabs saturated with suspensions of vaccinia virus, HSV, or VZV eliminated the infectivity of these agents; however, DNA was detectable in most autoclaved samples in dilutions of 10(-1) to 10(-4) by LightCycler PCR. All not autoclaved specimens were detected by culture (infectivity) except for VZV and, in most dilutions of 10(-1) to 10(-3), by assay of target DNA by LightCycler PCR. Similarly positive results were obtained for PCR assessment of sporulated B anthracis. CONCLUSIONS: Standard autoclaving procedures eliminated the infectivity of viruses (and B anthracis), but target DNA was often retained for detection by LightCycler PCR. Current recommendations indicate that the laboratory diagnosis of smallpox virus infection be performed only within Biosafety Level 4 facilities. We suggest that, in addition to the requirement for immediate coordination with public health officials, the federal government consider expanding the existing guidelines for processing these specimens to encourage immediate collection, autoclaving, and testing by LightCycler PCR to differentiate smallpox virus from other dermal pathogens such as HSV and VZV by specific qualified laboratories.
机译：目的：确定牛痘病毒，单纯疱疹病毒（HSV），水痘带状疱疹病毒（VZV）和炭疽芽孢杆菌的高压灭菌悬浮液是否能使这些药物失去感染力，但可以通过LightCycler聚合酶链反应（PCR）检测目标DNA。材料和方法：将拭子插入含有牛痘病毒，HSV，VZV或10的单个悬浮液的连续10倍稀释液（10（-1）至10（-5）; 500 microL;每个稀释液6个样品）的试管中（8）炭疽杆菌的菌落形成单位（2个样品）。一半的样品被高压灭菌，其余的则没有。将每个未高压灭菌的样品的等分试样用作感染性的对照。结果：用牛痘病毒，HSV或VZV悬液饱和的高压灭菌棉签消除了这些药物的传染性。但是，通过LightCycler PCR，可以在大多数高压灭菌的样品中以10（-1）至10（-4）的稀释度检测到DNA。通过培养（感染性）检测所有未高压灭菌的样品（VZV除外），并在大多数稀释度10（-1）至10（-3）中，通过LightCycler PCR分析目标DNA。类似地，阳性结果也可用于产孢炭疽热的PCR评估。结论：标准高压灭菌程序消除了病毒（和炭疽杆菌）的传染性，但目标DNA经常被保留以供LightCycler PCR检测。当前的建议表明，只能在生物安全4级设施内对天花病毒感染进行实验室诊断。我们建议，除了要求与公共卫生官员立即协调外，联邦政府还考虑扩大现有的处理这些标本的准则，以鼓励立即收集，高压灭菌和通过LightCycler PCR进行检测，以将天花病毒与其他皮肤病原体区分开来。由特定的合格实验室作为HSV和VZV。

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