Real Time PCR and Culture-Based Virus Isolation Test in Clinically Recovered Patients: Is the Subject Still Infectious for SARS-CoV2?
Real Time PCR and Culture-Based Virus Isolation Test in Clinically Recovered Patients: Is the Subject Still Infectious for SARS-CoV2?
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DOI：
10.3390/jcm10020309
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发表时间：
2021-01-15
影响因子：
3.9
通讯作者：
Foschino Barbaro MP
中科院分区：
文献类型：
--
作者：
Manzulli V;Scioscia G;Giganti G;Capobianchi MR;Lacedonia D;Pace L;Cipolletta D;Tondo P;De Nittis R;Rondinone V;Serrecchia L;Parisi A;Galante D;Lo Caputo S;Santantonio TA;Moschetta D;Dattoli V;Fasanella A;Foschino Barbaro MP
背景资料。新冠肺炎病的表现形式多种多样，从完全没有症状到致命，既是一个临床问题，也是一个公共卫生问题。到目前为止，住院患者的出院标准是基于实时逆转录聚合酶链式反应(RT-PCR)的阴性结果，但病毒片段的持久性可能会超过完整病毒的持久性数周。我们研究的目的是验证因新冠肺炎住院的临床康复但鼻咽拭子仍阳性的患者在病毒培养中的病毒清除情况。方法：研究方法。这项研究是在鼻咽拭子RT-PCR阳性的住院患者中进行的。患者从无症状到严重病例，在无症状患者第14天或症状缓解后至少3天进行鼻咽对照拭子。RT-PCR阳性标本被送往生物安全3级实验室进行病毒培养。结果。我们对84例住院患者进行了RT-PCR和高敏感性体外培养的联合分析。平均年龄46±20.29岁，40.5%的受试者经放射学确诊为肺炎，平均PaO2为72.35±12.12，P/F值为315±83.15。第一次拭子中N基因的CT值低于对照样本(p<；0.001)。83名患者的样本在病毒培养中呈阴性，相应培养上清液的RT-PCR总是证实没有病毒生长。结论。我们的初步结果表明，临床康复至少三天的患者在病毒培养中显示病毒清除，推测他们继续不具传染性。
Background. The highly variable manifestation of the COVID-19 disease, from completely asymptomatic to fatal, is both a clinical and a public health issue. The criteria for discharge of hospitalized patients have been based so far on the negative result of Real-Time Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) tests, but the persistence of viral fragments may exceed that of the integral virus by weeks. The aim of our study was to verify the clearance of the virus at viral culture in patients hospitalized for COVID-19 that have clinically recovered but are still positive on nasopharyngeal swab. Methods. The study was conducted in hospitalized patients with positive RT-PCR on nasopharyngeal swab. Patients included were from asymptomatic to severe cases and performed nasopharyngeal control swabbing on day 14 for asymptomatic patient or at least three days after remission of symptoms. RT-PCR positive specimens were sent to a biosafety level 3 laboratory for viral culture. Results. We performed a combined analysis of RT-PCR and a highly sensitive in vitro culture from 84 samples of hospitalized patients. The average age was 46 ± 20.29, and 40.5% of the subjects had radiologically confirmed pneumonia, with average PaO2 of 72.35 ± 12.12and P/F ratio of 315 ± 83.15. Ct values for the N gene were lower in the first swab than in the control one (p < 0.001). The samples from 83 patients were negative at viral culture, and RT-PCR on the respective supernatants always confirmed the absence of viral growth. Conclusions. Our preliminary results demonstrate that patients clinically recovered for at least three days show the viral clearance at viral culture, and presumably they continued to not be contagious.
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