Ng Enders K O, Hui David S, Chan K C Allen, Hung Emily C W, Chiu Rossa W K, Lee Nelson, Wu Alan, Chim Stephen S C, Tong Yu K, Sung Joseph J Y, Tam John S, Lo Y M Dennis
Department of Chemical Pathology, The Chinese University of Hong Kong, Prince of Wales Hospital, Hong Kong Special Administrative Region, China.
Clin Chem. 2003 Dec;49(12):1976-80. doi: 10.1373/clinchem.2003.024125.
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.
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.
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).
Plasma/serum SARS-CoV quantification represents a potentially useful early diagnostic and prognostic tool for SARS.
一种用于严重急性呼吸综合征（SARS）的早期诊断工具的可用性将具有重大的公共卫生意义。我们研究了在SARS早期阶段能否在血清和血浆样本中检测到SARS冠状病毒（SARS-CoV），并研究了这种方法潜在的预后意义。
我们开发了两种实时定量逆转录聚合酶链反应（RT-PCR）检测方法，一种用于病毒基因组的聚合酶区域，另一种用于核衣壳区域，以测量SARS患者血清/血浆样本中SARS-CoV RNA的浓度。在入院当天以及发热开始后第7天和第14天，从12例确诊的SARS患者中采集血浆样本。还在入院当天从23例确诊的SARS患者中采集血清样本，其中11例随后需要重症监护。从血浆/血清样本中提取病毒RNA。提取的RNA通过RT-PCR检测进行分析。
在发病第一周，针对聚合酶区域的RT-PCR系统在50%的SARS患者血浆样本和78%的血清样本中检测到SARS-CoV RNA。发热开始后第14天，血浆检测率降至25%。在住院期间需要和不需要入住重症监护病房的患者中，血清SARS-CoV浓度中位数分别为5800和140拷贝/毫升（Mann-Whitney检验，P<0.005）。针对核衣壳区域的RT-PCR系统证实了这些数据，其检测率甚至更高，为87%。两种RT-PCR系统获得的结果之间相关性很高（Pearson相关分析，r=0.998；P<0.001）。
血浆/血清SARS-CoV定量检测是一种对SARS潜在有用的早期诊断和预后工具。
