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将定量逆转录聚合酶链反应用于严重急性呼吸综合征冠状病毒2单引导核糖核酸以制定治疗方案:10例患者的初步报告

Use of qRT-PCR for SARS-CoV-2 sgRNA leader for the therapeutic plan: a preliminary report on 10 patients.

作者信息

Petcharat Sininat, Virojanapirom Phatthamon, Putcharoen Opass, Wacharapluesadee Supaporn, Hemachudha Thiravat

机构信息

Thai Red Cross Emerging Infectious Diseases Health Science Centre, King Chulalongkorn Memorial Hospital, Bangkok, Thailand.

Thai Red Cross Emerging Infectious Diseases Clinical Center, King Chulalongkorn Memorial Hospital, Bangkok, Thailand.

出版信息

J Infect Dev Ctries. 2022 Apr 30;16(4):604-607. doi: 10.3855/jidc.14852.

Abstract

INTRODUCTION

Duration of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) shedding is important for infection control. The presence of SARS-CoV-2 subgenomic RNA (sgRNA) leader indicates that the virus is replicative. This study examined the shedding duration of SARS-CoV-2 sgRNA leader and genomic RNA (gRNA) in diverse respiratory specimens.

METHODOLOGY

One hundred and eleven respiratory specimens collected sequentially from 10 COVID-19 patients with real-time RT-PCR SARS-CoV-2 orf1ab gene confirmed positive admitted to King Chulalongkorn Memorial Hospital were examined for SARS-CoV-2 E sgRNA leader and E gRNA by using Real-time reverse transcription PCR (qRT-PCR). These specimens included nasopharyngeal swab and throat swabs, nasal swab and throat swabs, sputum, and endotracheal aspirate, and were collected from the first day of admission until the time of orf1ab real-time RT-PCR negative of at least 2-4 consecutive days.

RESULTS

E sgRNA leader could only be detectable in specimens with ≥ 1E+05 virus E gene copies per ml within the first 15 days after hospitalization. SARS-CoV-2 sgRNA leader was undetectable from one to 15 days earlier than that of gRNA in all patients. Re-shedding of sgRNA was evident in 2 cases, both on a single occasion after being undetectable for 3-10 days.

CONCLUSIONS

Assessment of the presence of sgRNA leader may be useful for therapeutic planning.

摘要

引言

严重急性呼吸综合征冠状病毒2(SARS-CoV-2)的排毒持续时间对于感染控制很重要。SARS-CoV-2亚基因组RNA(sgRNA)前导序列的存在表明该病毒正在复制。本研究检测了SARS-CoV-2 sgRNA前导序列和基因组RNA(gRNA)在不同呼吸道标本中的排毒持续时间。

方法

对朱拉隆功国王纪念医院收治的10例经实时逆转录聚合酶链反应(RT-PCR)检测SARS-CoV-2 orf1ab基因确诊为阳性的COVID-19患者,连续采集111份呼吸道标本,采用实时逆转录聚合酶链反应(qRT-PCR)检测SARS-CoV-2 E sgRNA前导序列和E gRNA。这些标本包括鼻咽拭子、咽喉拭子、鼻腔拭子、痰液和气管内吸出物,从入院第一天开始采集,直至orf1ab实时RT-PCR至少连续2-4天呈阴性。

结果

E sgRNA前导序列仅在住院后前15天内每毫升病毒E基因拷贝数≥1E+05的标本中可检测到。在所有患者中,SARS-CoV-2 sgRNA前导序列比gRNA早1至15天检测不到。2例患者出现sgRNA再次排毒,均在检测不到3-10天后单次出现。

结论

评估sgRNA前导序列的存在可能有助于治疗方案的制定。

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