Effects of fixative type and fixation time on the detection of Maedi Visna virus by PCR and immunohistochemistry in paraffin-embedded ovine lung samples
Effects of fixative type and fixation time on the detection of Maedi Visna virus by PCR and immunohistochemistry in paraffin-embedded ovine lung samples
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DOI：
10.1016/j.jviromet.2006.07.009
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发表时间：
2006-11-01
影响因子：
3.1
通讯作者：
Perez, Valentin
中科院分区：
文献类型：
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作者：
Benavides, Julio;Garcia-Pariente, Carlos;Perez, Valentin
在可疑病例中，由 Maedi Visna 病毒（MVV）（一种绵羊慢性多系统慢病毒病）引起的病变的组织病理学诊断需要通过组织中 MVV 的表现来确认。在来自五只血清学阳性羊的不同程度病变的肺样本中，评估了固定剂和固定时间对石蜡包埋组织中免疫组织化学 (IHC) 和 PCR 检测 MVV 的影响。将样品固定在 10% 中性缓冲福尔马林 (NBF)、博宁溶液 (BS) 和锌盐固定剂 (ZSF) 中，固定时间为 24 小时至 30 天。三种固定剂保留了组织的形态，尽管在 ZSF 固定的样品中，肺泡中脱落细胞的数量有所增加。无论使用 ZSF 和 NBF 固定剂固定的时间长短，组织都显示出相似程度的免疫标记。在 NBF 中固定最多 14 天、在 ZSF 中固定 30 天的样品中可以检测到 MVV 核酸。然而。在BS固定的组织中，固定4天后免疫染色很弱并且观察到非特异性信号。在这些样本中无法通过 PCR 获得原病毒 DNA 的扩增。 IHC 在所有绵羊中均检测到了病毒抗原，而一只患有轻度病变的绵羊则通过 PCR 检测始终呈阴性。 (c) 2006 Elsevier B.V. 保留所有权利。
In doubtful cases, the histopathological diagnosis of lesions induced by Maedi Visna virus (MVV), a chronic multisystemic lentiviral disease of sheep, needs to be confirmed by the demonstration of MVV in the tissues. The influence of fixatives and the duration of fixation on the detection of MVV by immunohistochemistry (IHC) and PCR in paraffin-embedded tissues was assessed in lung samples with lesions in different degree, from five sheep serologically positive. Samples were fixed in 10% neutral buffered formalin (NBF), Bonin's solution (BS) and a zinc salts-based fixative (ZSF), for different periods of time between 24 h and 30 days. The three fixatives preserved the morphology of the tissues, although in ZSF-fixed samples an increase in the number of desquamated cells was seen in the alveoli. Tissues showed a similar degree of immunolabelling, irrespective of the duration of fixation using ZSF and NBF fixatives. MVV nucleic acids could be detected in samples fixed up to 14 days in NBF and 30 days in ZSF. However. in BS fixed tissues, immunostaining was weak and non-specific signals were observed after 4 days of fixation. Amplification of proviral DNA could not be obtained by PCR in these samples. IHC detected viral antigens in all sheep whereas one sheep with mild lesions was always negative by PCR. (c) 2006 Elsevier B.V. All rights reserved.