Nucleic acid detection

核酸检测
  • 文章类型: Journal Article
    探讨预测无症状和轻度COVID-19患者核酸阴性转化时间和ORF1ab基因CT值变化的影响因素。
    选取2022年4月3日至4月23日上海市移动客舱医院收治的73,456例无症状且轻度COVID-19患者作为研究对象。流行病学,临床,并收集了实验室数据。进行相关性分析。
    在<18岁和≥65岁的患者中,COVID-19疫苗可以缩短核酸阴性转化时间,这反映在较低的中位数或75%的分位数(P<0.001,P<0.05)。在患有潜在疾病的患者中,加强接种和完全接种的阴性核酸转化时间均低于未接种组(P<0.001,P<0.05)。≤18岁或>65岁的患者,合并症患者和有症状的患者,与18-65岁的患者相比,没有合并症的患者和没有症状的患者,接种和未接种的患者的CT值上升率差异较大(P<0.05)。
    无症状和轻度COVID-19患者的核酸转化为阴性的时间受年龄的影响,合并症,和第一核酸CT值。接种疫苗可以缩短老年人的核酸阴性转化时间,那些有并发症或症状的人。老年患者的疫苗接种不会增加症状的风险。
    UNASSIGNED: To explore the influencing factors to predict the negative nucleic acid conversion time and ORF1ab gene CT value changes in patients with asymptomatic and mild COVID-19.
    UNASSIGNED: A total of 73,456 patients with asymptomatic and mild COVID-19 admitted to the Mobile Cabin Hospital in Shanghai from April 3 to April 23, 2022 were selected as the research objects. Epidemiological, clinical, and laboratory data were collected. Correlation analysis was performed.
    UNASSIGNED: In patients <18 years old and ≥65 years old, COVID-19 vaccine can shorten the negative nucleic acid conversion time, which is reflected in the lower median or 75% quantile (P<0.001, P<0.05). In patients with underlying diseases, the negative nucleic acid conversion time of booster vaccination and complete vaccination was lower than that of non-vaccinated group (P<0.001, P<0.05). In patients ≤18 years of age or >65 years of age, patients with comorbidity and patients with symptoms, compared with patients 18-65 years of age, patients without comorbidity and patients without symptoms, there was a greater difference in the rate of rise of CT values between vaccinated and unvaccinated patients (P<0.05).
    UNASSIGNED: The time of nucleic acid conversion to negative in patients with asymptomatic and mild COVID-19 is affected by age, comorbidity, and first nucleic acid CT value. Vaccination could shorten the negative nucleic acid conversion time of the older population, those with complications or symptoms. The vaccination of older patients does not increase the risk of symptoms.
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  • 文章类型: Journal Article
    COVID-19疫情的爆发使各种快速检测方法得以建立和应用。建立一种快速、准确的肠道病毒检测方法尤为重要,这将有利于临床诊断,疫情防控,和及时的可追溯性。通过建立超快速逆转录-聚合酶链反应(RT-PCR)设备,本研究旨在评价基于超快实时荧光RT-PCR技术的肠道病毒核酸检测方法的敏感性和特异性。共抽取61例,然后被运输和保存。核酸提取后,用广州大安基因公司生产的肠道病毒核酸检测试剂盒和本研究建立的超快速RT-PCR设备系统检测同一样品的核酸。ABI7500Fast和Ahram生物系统S1快速设备用于扩增检测。如果样品在FAM通道中具有S形扩增曲线并且Ct值≤40.00,则结果为阳性。敏感性,精度,验证了检测方法的准确性。本研究建立了一种新的检测方法,可在24min内实现肠道病毒核酸的检测。该方法的灵敏度检测极限为1.0×102拷贝/ml。重复检测的变异系数高,中等,低浓度样品为2.644%,1.674%,和4.281%,分别,具有良好的检测重复性。此外,61例可疑样本中,超快速RT-PCR检测方法阳性29例,与常规荧光RT-PCR方法一致。建立的快速检测方法可以大大缩短提供检测报告的时间,可以大大提高诊断和治疗的效率。
    The outbreak of COVID-19 epidemic has enabled the establishment and application of various rapid detection methods. It is particularly important to establish a fast and accurate detection method for enterovirus, which will be beneficial for clinical diagnosis, epidemic prevention and control, and timely traceability. Through establishing an ultra-fast reverse transcription-polymerase chain reaction (RT-PCR) equipment, this study aimed to evaluate the sensitivity and specificity of the testing method of enterovirus nucleic acids based on ultra-fast real-time fluorescence RT-PCR technology. A total of 61 cases were sampled, which were then transported and preserved. After the nucleic acid extraction, the nucleic acids of the same sample were tested with the enterovirus nucleic acid detection kit produced by Guangzhou Da An Gene Company and the ultra-fast RT-PCR equipment system established in this study. ABI7500Fast and Ahram biosystems S1 fast equipment were used for amplification detection. If the sample had an S-shaped amplification curve in the FAM channel and the Ct value ≤40.00, the result was positive. The sensitivity, precision, and accuracy of the detection method were then verified. This study established a novel testing method to achieve enterovirus nucleic acid detection within 24 min. The sensitivity detection limit of the method was 1.0 × 102 copies/ml. The coefficients of variation for repeated detection of the high, medium, and low concentration samples were 2.644%, 1.674%, and 4.281%, respectively, with good detection repeatability. In addition, a total of 29 cases were positive by the ultra-fast RT-PCR detection method in 61 suspected samples, which was consistent with the conventional fluorescent RT-PCR method. The established rapid detection method can greatly shorten the time for providing a detection report, which may greatly improve the efficiency of diagnosis and treatment.
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  • 文章类型: Journal Article
    Nucleic acid detection technique has good sensitivity and specificity and is widely used in in vitro diagnosis, animal and plant commodity quarantine, forensic identification, and other fields. However, it is susceptible to carryover contamination during the operation and leads to false-positive results, which seriously affects the detection accuracy. Therefore, finding an effective solution to prevent and eliminate nucleic acid carryover contamination has become particularly urgent. This study compared several different methods for removing nucleic acid contamination and confirmed that sodium hypochlorite solution and PCRguard reagent could effectively eliminate nucleic acid carryover in the liquid and on surfaces of different materials. Besides, the combination of sodium hypochlorite solution and PCRguard can solve the nucleic acid aerosol contamination. This study proposes solutions for the routine prevention of carryover contamination and removal of aerosol that has occurred in molecular diagnostic laboratories.
    核酸检测因具有良好的灵敏度和特异性而被广泛应用于体外诊断、动植物商品检疫、法医鉴定等领域。然而操作过程中易受到核酸污染导致的假阳性结果,严重影响了检测准确性。因此寻找一种有效的防止和清除核酸污染的方案对于实验室正常运转及保障检测结果的可靠性具有重要意义。文中比较了几种不同清除核酸污染的方法,确认了84消毒液和PCRguard试剂可有效清除液体中及不同材质表面的核酸污染。另外,84消毒液和PCRguard试剂配合使用,可很好地解决核酸气溶胶污染。研究结果对于分子诊断实验室日常预防核酸污染及清除已发生的气溶胶污染提出了解决方案。.
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