Quantitative analysis and prognostic implication of SARS coronavirus RNA in the plasma and serum of patients with severe acute respiratory syndrome
Quantitative analysis and prognostic implication of SARS coronavirus RNA in the plasma and serum of patients with severe acute respiratory syndrome
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DOI：
10.1373/clinchem.2003.024125
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发表时间：
2003-12-01
影响因子：
9.3
通讯作者：
Lo, YMD
中科院分区：
文献类型：
--
作者：
Ng, EKO;Hui, DS;Lo, YMD
背景：早期诊断工具的可用性。严重急性呼吸系统综合症（SARS）将对公共卫生产生重大影响。我们调查了SARS冠状病毒（SARS-CoV）是否可以在SARS早期的血清和血浆样本中检测到，并研究了可能性。方法：我们开发了两种实时定量逆转录-PCR（RT-PCR）检测，一种用于聚合酶，另一种用于病毒基因组的核衣壳区域，用于测量SARS患者血清/血浆样本中SARS冠状病毒RNA的浓度。12名确诊SARS患者在入院当天以及发烧后第7天和第14天采集血浆样本。此外，我们亦于23名确诊SARS病人入院当日抽取他们的血清样本，其中11名病人其后需要深切治疗。从血浆/血清样品中提取病毒RNA。结果：SARS患者发病第1周内，SARS冠状病毒聚合酶区的RT-PCR检测率为50%，血清标本的检出率为78%。发热后第14天血浆检出率降至25%。在住院期间需要和不需要重症监护病房的患者中位血清SARS-CoV浓度分别为5800和140拷贝/mL（Mann-Whitney检验，P < 0.005）。这些数据被核衣壳区的RT-PCR系统证实，其显示出更高的检测率为87%。结论：血浆/血清SARS-CoV定量检测对SARS的早期诊断和预后判断具有重要意义。(C)2003年美国临床化学协会。
Background: The availability of an early diagnostic tool. for severe acute respiratory syndrome (SARS) would have major public health implications. We investigated whether the SARS coronavirus (SARS-CoV) can be detected in serum and plasma samples during the early stages of SARS and studied the potential. prognostic implications of such an approach.Methods: We developed two real-time quantitative reverse transcription-PCR (RT-PCR) assays, one for the polymerase and the other for the nucleocapsid region of the virus genome, for measuring the concentration of SARS-CoV RNA in serum/plasma samples from SARS patients. Plasma samples were obtained from 12 confirmed SARS patients on the day of hospital admission, as well as on days 7 and 14 after fever onset. Serum samples were-also obtained from 23 confirmed SARS patients on the day of hospital admission, 11 of whom subsequently required intensive care. Viral RNA was extracted from the plasma/serum samples. The extracted RNA was subjected to analysis by the RT-PCR assays.Results: The RT-PCR system for the polymerase region detected SARS-CoV RNA in 50% of plasma and 78% of serum samples from SARS patients during the first week of illness. The detection rates for plasma dropped to 25% at day 14 after fever onset. The median serum SARS-CoV concentrations in patients who required and did not require intensive care unit admission during the course of hospitalization were 5800 and 140 copies/mL, respectively (Mann-Whitney test, P < 0.005). These data were confirmed by the RT-PCR system for the nucleocapsid region, which showed an even higher detection rate of 87%. The correlation between the results obtained by the two RT-PCR systems was high (Pearson correlation analysis, r = 0.998; P < 0.001).Conclusion: Plasma/serum SARS-CoV quantification represents a potentially useful early diagnostic and prognostic tool for SARS. (C) 2003 American Association for Clinical Chemistry.