respiratory pathogens

呼吸道病原体
  • 文章类型: Journal Article
    在由COVID-19等呼吸道病原体引起的大流行期间,在加拿大NICU中共同创建父母在场实践建议。
    建议是通过证据提出的,context,Delphi和值和首选项方法。对于Delphi1和2,参与者分别对50个项目和20个项目进行了评分,从1(重要性非常低)到5(非常高)。为了确定共识,在排名靠前的项目的价值和偏好框架内提出并讨论了益处和危害的证据和背景。80%或更多的协议被视为共识。
    经过两轮德尔菲(n=59名参与者),确定了13项评级重要性最高的建议。共识建议包括6项强有力的建议(父母作为基本照顾者,提供皮肤与皮肤的接触,直接或母亲自己表达的牛奶喂养,参加医疗查房,心理健康和社会心理服务,并将父母伙伴纳入大流行应对计划)和7项有条件建议(提供动手护理任务,提供触摸,两位家长同时在场,食物和饮料的获取,使用通信设备,以及当面获得医疗查房、心理健康和社会心理服务)。
    这些建议可以指导机构制定在COVID-19等呼吸道病原体引起的大流行期间父母存在的策略。
    UNASSIGNED: To co-create parental presence practice recommendations across Canadian NICUs during pandemics caused by respiratory pathogens such as COVID-19.
    UNASSIGNED: Recommendations were developed through evidence, context, Delphi and Values and Preferences methods. For Delphi 1 and 2, participants rated 50 items and 20 items respectively on a scale from 1 (very low importance) to 5 (very high). To determine consensus, evidence and context of benefits and harms were presented and discussed within the Values and Preference framework for the top-ranked items. An agreement of 80% or more was deemed consensus.
    UNASSIGNED: After two Delphi rounds (n = 59 participants), 13 recommendations with the highest rated importance were identified. Consensus recommendations included 6 strong recommendations (parents as essential caregivers, providing skin-to-skin contact, direct or mothers\' own expressed milk feeding, attending medical rounds, mental health and psychosocial services access, and inclusion of parent partners in pandemic response planning) and 7 conditional recommendations (providing hands-on care tasks, providing touch, two parents present at the same time, food and drink access, use of communication devices, and in-person access to medical rounds and mental health and psychosocial services).
    UNASSIGNED: These recommendations can guide institutions in developing strategies for parental presence during pandemics caused by respiratory pathogens like COVID-19.
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  • 文章类型: Journal Article
    呼吸道病毒感染,包括呼吸道合胞病毒(RSV),副流感病毒和A型和B型流感病毒,可能会有严重的结果。细菌感染经常跟随病毒感染,和流感或其他病毒流行定期有更高的死亡率由继发性细菌性肺炎。大多数继发性细菌感染可通过激活细胞受体以操纵嗜中性粒细胞的脂肪酸介质引起肺免疫抑制,巨噬细胞,自然杀伤细胞,树突状细胞和其他肺免疫细胞。细菌感染诱导炎症介质的合成,包括前列腺素和白三烯,然后最终也是特别支持解决的调解员,包括脂蛋白,resolvins,protectinsandmaresins,通常可以解决炎症和免疫抑制。并发的病毒和继发细菌感染更危险,因为在继发细菌感染加重炎症和免疫抑制之前,病毒感染会引起炎症和免疫抑制。很有可能,继发性细菌性肺炎的死亡率较高是由压倒性的炎症和免疫抑制引起的,特别支持解决的调解员可能无法解决。
    Respiratory viral infections, including respiratory syncytial virus (RSV), parainfluenza viruses and type A and B influenza viruses, can have severe outcomes. Bacterial infections frequently follow viral infections, and influenza or other viral epidemics periodically have higher mortalities from secondary bacterial pneumonias. Most secondary bacterial infections can cause lung immunosuppression by fatty acid mediators which activate cellular receptors to manipulate neutrophils, macrophages, natural killer cells, dendritic cells and other lung immune cells. Bacterial infections induce synthesis of inflammatory mediators including prostaglandins and leukotrienes, then eventually also special pro-resolving mediators, including lipoxins, resolvins, protectins and maresins, which normally resolve inflammation and immunosuppression. Concurrent viral and secondary bacterial infections are more dangerous, because viral infections can cause inflammation and immunosuppression before the secondary bacterial infections worsen inflammation and immunosuppression. Plausibly, the higher mortalities of secondary bacterial pneumonias are caused by the overwhelming inflammation and immunosuppression, which the special pro-resolving mediators might not resolve.
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  • 文章类型: Journal Article
    在中国患有社区获得性肺炎(CAP)的成年人发病率很高。CAP是由多种病原体引起的;然而,通常缺乏针对病原体的临床症状。因此,缺乏准确微生物学诊断的患者接受经验性抗微生物药物治疗.
    我们收集了支气管肺泡灌洗液,以及湖北三家医院收治的650名成人CAP患者的临床和实验室数据,四川,和中国的浙江省。培养样本,并使用实时逆转录qPCR(RT-qPCR)测定法测试42种呼吸道细菌和病毒的存在。对CAP进行了地区调查,性别,以及感染或合并感染的年龄和模式。采用适合诊断的临床指南,我们回顾性评估了适当的病原体导向治疗,并将其与最初的经验性治疗进行了比较.
    我们的研究发现,21.38%(139/650)的患者被归类为严重CAP(S-CAP),男性患病率较高,老年人,在温暖的季节。35.53%(231/650)的病例检出细菌病原体。肺炎克雷伯菌,流感嗜血杆菌,金黄色葡萄球菌是不同人口统计学和地区最普遍的细菌。在48.76%(317/650)的患者中发现了病毒病原体,人类鼻病毒,巨细胞病毒是最常见的病毒。24.31%(158/650)的病例存在合并感染,病毒-细菌共感染是最常见的。与标准培养方法相比,RT-qPCR对关键病原体的检测率明显更高。通过允许18.30%(95/518)的患者降级,它显示了优化抗菌药物处方的潜力,其中减少过量抗生素的数量主要包括减少第二代或第三代头孢菌素的使用(5.79%,30/518)和β-内酰胺酶抑制剂组合。
    该研究强调了S-CAP的重大负担,特别是在特定的人口统计和季节。细菌和病毒病原体的流行,伴随着高感染率,强调需要全面的诊断方法。RT-qPCR检测是一种卓越的诊断工具,提供增强的病原体检测能力和促进更精确的抗菌治疗。这可以改善患者的预后,并有助于合理使用抗菌药物,解决日益增长的抗生素耐药性问题。
    UNASSIGNED: Adults with community-acquired pneumonia (CAP) in China suffer high morbidity. CAP is caused by a multitude of pathogens; however, pathogen-directed clinical symptoms are often lacking. Therefore, patients lacking an accurate microbiological diagnosis are administered with empirical antimicrobials.
    UNASSIGNED: We collected bronchoalveolar lavage fluid, as well as clinical and laboratory data from 650 adult patients with CAP admitted to three hospitals in Hubei, Sichuan, and Zhejiang provinces in China. Specimens were cultured and tested using real-time reverse transcription qPCR (RT-qPCR) assays for the presence of 42 respiratory bacteria and viruses. CAP was investigated with respect to regions, genders, and age and patterns of infections or co-infections. Employing clinical guidelines adapted for diagnosis, we assessed retrospectively the appropriate pathogen-directed therapy and compared it with the initial empirical therapies.
    UNASSIGNED: Our study identified that 21.38% (139/650) of the patients were classified as having Severe CAP (S-CAP), with a higher prevalence among males, older adults, and during the warm season. Bacterial pathogens were detected in 35.53% (231/650) of cases. K. pneumoniae, H. influenzae, and S. aureus were the most prevalent bacteria across different demographics and regions. Viral pathogens were found in 48.76% (317/650) of patients Epstein-Barr, Human rhinovirus, and Cytomegalovirus were the most common viruses. Co-infections were present in 24.31% (158/650) of cases, with viral-bacterial co-infections being the most frequent. The RT-qPCR demonstrated significantly higher detection rates for key pathogens compared to standard culture methods. It showed potential in optimizing antimicrobial prescriptions by allowing for de-escalation in 18.30% (95/518) of patients, among which reducing the number of excessive antibiotics mainly comprised decreasing the use of 2nd or 3rd generation cephalosporins (5.79%, 30/518) and β-lactamase inhibitor combinations.
    UNASSIGNED: The study highlights the significant burden of S-CAP, particularly among specific demographics and seasons. The prevalence of bacterial and viral pathogens, along with the high rate of co-infections, emphasizes the need for comprehensive diagnostic approaches. The RT-qPCR assays emerge as a superior diagnostic tool, offering enhanced pathogen detection capabilities and facilitating more precise antimicrobial therapy. This could lead to improved patient outcomes and contribute to the rational use of antimicrobials, addressing the growing concern of antibiotic resistance.
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  • 文章类型: Journal Article
    尽管共同感染的流行和超过50个病毒和46个细菌病原体与猪疾病的关联,对它们同时发生的情况知之甚少,特别是在健康计划到位的商业养猪环境中。为了解决这个知识差距,本研究旨在使用PorkMultiPath™(PMP1和PMP2,分别为呼吸道和肠道)技术评估猪群中呼吸道和肠道病原体的病原体阈值,在单个反应中同时检测多种病原体,具有高灵敏度和特异性。在这项研究中,最常见的呼吸道病原体,支原体hyrohinis,多杀性巴氏杆菌,通过tiamulin的策略治疗,PMP1检测到的副猪嗜血杆菌在苗圃阶段得到了有效控制。即使主要的呼吸道发病率减少了,记录的咳嗽和打喷嚏率与副猪嗜血杆菌和猪嗜血杆菌的水平有关,设置为1356和1275个拷贝/反应,分别。此外,确定的共感染模式之一表明副猪嗜血杆菌和猪鼻分枝杆菌在样本和围栏水平的发生之间有很强的关系,突出了一起检测这两种病原体的可能性很高。用肠面板PMP2进行的测试表明,在苗圃早期阶段最常见的毒力因子是毒素的大肠杆菌基因-ST1,ST2和菌毛-F4和F18。此外,在苗圃阶段经常观察到轮状病毒B和C的共感染,这两个标记之间具有很强的正相关性。此外,几个标记的水平,即大肠杆菌F4,F5,F18,LT,ST1和ST2与猪群体中更高的患病可能性相关。此外,发现在苗圃和种植者阶段出现短螺旋体与腹泻风险增加有关,设定的阈值在约500个拷贝/反应。尽管多种病原体的同时检测在养猪业中尚未广泛使用,它在捕捉共感染的多样性和相互作用方面具有显著优势。使用PorkMultiPath™测试汇集的样品具有成本效益,可定期监测猪群的健康状况。
    Despite the prevalence of co-infections and the association of over 50 viral and 46 bacterial pathogens with pig diseases, little is known about their simultaneous occurrence, particularly in commercial pig farming environments where health programs are in place. To address this knowledge gap, this study aimed to evaluate the pathogen threshold of respiratory and enteric pathogens in pig herds using the Pork MultiPath™ (PMP1 and PMP2, respiratory and enteric respectively) technology, which detects multiple pathogens simultaneously in a single reaction with high sensitivity and specificity. In this study the most prevalent respiratory pathogens, Mycoplasma hyrohinis, Pasteurella multocida, and Haemophilus parasuis detected by PMP1 were effectively controlled during the nursery stage through strategic treatment with tiamulin. Even though the major respiratory incidences were reduced, the recorded coughing and sneezing rates were associated with the levels of H. parasuis and M. hyrohinis, which were set at 1356 and 1275 copies/reaction, respectively. In addition, one of the identified co-infection patterns indicated a strong relationship between the occurrence of H. parasuis and M. hyorhinis at the sample and pen levels, highlighting the high likelihood of detecting these two pathogens together. Testing with enteric panel PMP2 revealed that the most frequently detected virulence factors during the early nursery stage were Escherichia coli genes for toxins - ST1, ST2, and fimbriae - F4 and F18. Moreover, a co-infection with Rotavirus B and C was often observed during the nursery stage, and a strong positive correlation between these two markers has been identified. Additionally, the levels of several markers, namely E. coli F4, F5, F18, LT, ST1, and ST2, have been associated with a higher likelihood of sickness in pig populations. In addition, the onset of Brachyspira pilosicoli during the nursery and grower stages was found to be associated with an increased risk of diarrhoea, with a set threshold at around 500 copies/reaction. Although simultaneous detection of multiple pathogens is not yet widely used in the pig industry, it offers a significant advantage in capturing the diversity and interactions of co-infections. Testing pooled samples with Pork MultiPath™ is cost-effective and practical to regularly monitor the health status of pig populations.
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  • 文章类型: Journal Article
    COVID-19已成为我们这个时代最重要的全球健康问题。病原体,SARS-CoV-2对易感人群的下呼吸道造成广泛损害,导致肺部损伤和死亡.感染COVID-19的患者也容易感染呼吸道病原体,如铜绿假单胞菌,金黄色葡萄球菌,肺炎克雷伯菌,和大肠杆菌。在某些情况下,这些呼吸道病原体具有多重耐药性,可导致患者感染危及生命。由于现有的抗生素对这些抗生素耐药细菌无效,迫切需要注意开发新的和有效的治疗剂来对抗抗微生物细菌。或者,可以探索新的治疗策略来增强现有抗微生物剂的抗微生物效果,比如抗生素。将天然化合物与现有的抗菌剂一起添加以增加其抗微生物活性是应对COVID-19和抗菌素耐药性不断上升的威胁的最合适和有希望的选择之一。天然化合物通常被认为是安全的,甚至可以减少药物和药物的副作用。鉴于这些优势,本综述总结了一些将天然化合物与抗生素和抗病毒药物结合起来以增加这些药物的抗菌潜力的研究。这项研究可以帮助研究人员比较和理解已经存在的数据,以设计新的研究来开发针对COVID-19的抗菌药物。
    COVID-19 has emerged as the most significant global health issue of our time. The causative agent, SARS-CoV-2, causes extensive damage to the lower respiratory tract in susceptible populations, leading to lung damage and death. COVID-19-infected patients are also prone to respiratory pathogens such as Pseudomonas aeruginosa, Staphylococcus aureus, Klebsiella pneumoniae, and E. coli. In some cases, these respiratory pathogens are multidrug-resistant and cause life-threatening infections in patients. Since the existing antibiotics are ineffective against these antibiotic-resistant bacteria, urgent attention is required to develop new and effective therapeutic agents to combat antimicrobial-resistant bacteria. Alternatively, novel therapeutic strategies can be explored to enhance the antimicrobial effects of the existing antimicrobial agents, such as antibiotics. Adding natural compounds with existing antimicrobial agents to increase their antimicrobial activity is one of the most suitable and promising options to combat the rising threat of both COVID-19 and antimicrobial resistance. Natural compounds are generally considered safe and may even reduce the side effects of drugs and medicines. In light of such advantages, the current review summarized some of the studies that have combined natural compounds with antibiotics and antiviral to increase the antimicrobial potential of these drugs. This study can help researchers compare and understand already existing data to design new studies to develop antimicrobial agents against COVID-19.
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  • 文章类型: Journal Article
    本研究旨在探讨宏基因组下一代测序(mNGS)在肺弥漫性渗出性病变中的临床应用价值。
    从2014年1月1日至2021年11月31日,福建省立医院收治的136例胸部影像学表现为肺弥漫性渗出性病变的患者纳入研究;其中,77例患者行mNGS病原体检测。根据病原体检测结果和临床诊断,患者分为感染组(IG)和非感染组(NIG).比较了mNGS技术和传统培养方法的诊断效能。同时,59名临床鉴定为具有感染性肺弥漫性渗出性病变但未接受mNGS测试的患者被指定为非NGS感染组(非IG)。对IG和非IG患者进行了回顾性队列研究,30天全因死亡率终点用于随访。
    与常规培养方法相比,mNGS的灵敏度提高了约35%(80.0%vs45.5%,P<0.001),特异性无显著差异(77.3%vs95.5%,P=0.185)。在接触抗生素的情况下,mNGS检测的阳性率明显高于传统培养方法,表明mNGS受抗生素暴露的影响较小(P<0.05)。30天内,IG与非IG患者的全因死亡率分别为14.55%和37.29%,分别为(P<0.05)。在进行COX回归分析以调整混杂因素后,分析显示,CURB-65评分≥3分(HR=3.348,P=0.001)和存在心血管疾病(HR=2.473,P=0.026)是这些患者的独立危险因素.相反,mNGS检测(HR=0.368,P=0.017)是一个独立的保护因素。
    mNGS技术可以更轻松地查明肺部弥漫性感染性渗出性病变的原因,而不会受到抗生素的太多干扰,帮助医生尽早发现和诊断这些问题,从而在帮助他们为患者决定最佳治疗方法方面发挥关键作用。这样的结论可能有偏见,因为缺乏血清学检测和PCR等其他常规诊断技术的完整结果,传统方法的性能可能被低估。
    UNASSIGNED: This study aims to investigate the clinical application value of Metagenome Next-Generation Sequencing (mNGS) for pulmonary diffuse exudative lesions.
    UNASSIGNED: From January 1, 2014, to November 31, 2021, 136 cases with chest radiologic presentations of pulmonary diffuse exudative lesions admitted to Fujian Provincial Hospital were included in the study; of those, 77 patients underwent mNGS pathogen detection. Based on the pathogen detection outcomes and clinical diagnoses, patients were categorized into an infection group (IG) and a non-infection group (NIG). A comparison was made between the diagnostic efficacy of the mNGS technique and traditional culture methods. Meanwhile, 59 patients clinically identified as having infectious pulmonary diffuse exudative lesions but who did not receive mNGS testing were designated as the non-NGS infection group (non-IG). A retrospective cohort study was conducted on patients in both the IG and non-IG, with a 30-day all-cause mortality endpoint used for follow-up.
    UNASSIGNED: When compared to conventional culture methods, mNGS demonstrated an approximate 35% increase in sensitivity (80.0% vs 45.5%, P<0.001), without significant disparity in specificity (77.3% vs 95.5%, P=0.185). Under antibiotic exposure, the positivity rate detected by mNGS was notably higher than that by traditional culture methods, indicating that mNGS is less affected by exposure to antibiotics (P<0.05). Within 30 days, the all-cause mortality rate for patients in the IG versus the non-IG was 14.55% and 37.29%, respectively (P<0.05). Following a COX regression analysis to adjust for confounding factors, the analysis revealed that a CURB-65 score ≥3 points (HR=3.348, P=0.001) and existing cardiovascular disease (HR=2.473, P=0.026) were independent risk factors for these patients. Conversely, mNGS testing (HR=0.368, P=0.017) proved to be an independent protective factor.
    UNASSIGNED: mNGS technology makes it easier to pinpoint the cause of pulmonary diffuse infectious exudative lesions without much interference from antibiotics, helping doctors spot and diagnose these issues early on, thereby playing a key role in helping them decide the best treatment approach for patients. Such conclusions may have a bias, as the performance of traditional methods might be underestimated due to the absence of complete results from other conventional diagnostic techniques like serological testing and PCR.
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  • 文章类型: Journal Article
    呼吸道病原体,如SARS-CoV-2和A/B流感,会导致易感个体的严重疾病。这项研究评估了一种新型的数字微流体护理点测试平台,旨在检测23种病原体,将其性能与传统的基于实验室的核酸测试进行比较。该平台集成了核酸提取和扩增过程,仅需2分钟的动手时间即可快速检测。性能测定表明,该平台对所评估的3种病毒的检测具有高灵敏度(87%-100%)和特异性(99%-100%)。此外,该平台可用于检测其他呼吸道病原体,帮助呼吸系统疾病的早期诊断,确定爆发或流行病的来源,遏制疾病的传播。
    Respiratory pathogens, such as SARS-CoV-2 and influenza A/B, can cause severe illnesses in susceptible individuals. This research evaluated a novel digital microfluidic point-of-care testing platform designed to detect 23 pathogens, comparing its performance to conventional laboratory-based nucleic acid tests. The platform integrates nucleic acid extraction and amplification processes for rapid detection with only 2 min of hands-on time. Performance assays demonstrated that the platform has high sensitivity (87 %-100 %) and specificity (99 %-100 %) for the detection of the evaluated 3 viruses. Additionally, the platform can be adapted for the detection of other respiratory pathogens, aiding in the early diagnosis of respiratory diseases, identifying the source of an outbreak or epidemic, and curbing the spread of the disease.
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  • 文章类型: Journal Article
    背景:人类接触模式是驱动呼吸道传染病传播的关键决定因素。然而,接触模式与季节性之间的关系以及它们与呼吸系统疾病季节性之间可能的关联尚待澄清。
    方法:我们使用通过在上海进行的基于横断面日记的接触调查收集的数据,调查了温度与人类接触模式之间的关联,中国,2017年12月24日至2018年5月30日。然后,我们根据得出的接触人数的季节性趋势开发了流感传播的隔室模型,并根据同期在上海收集的A(H1N1)pdm09流感数据进行了验证。
    结果:我们确定了接触数量与季节性温度趋势之间的显着反比关系,该趋势定义为温度数据的样条插值(p=0.003)。我们估计2017年12月平均每天有16.4次(95%PrI:15.1-17.5)接触,2018年1月增加到平均17.6次(95%PrI:16.5-19.3),然后在2018年5月下降到平均10.3次(95%PrI:9.4-10.8)。通过隔室模型获得的流感发病率估计符合观察到的流行病学数据。繁殖数量估计从12月的1.24(95%CI:1.21-1.27)增加到1月的1.34(95%CI:1.31-1.37)的峰值。季节结束时估计的中位感染发生率为27.4%(95%CI:23.7-30.5%)。
    结论:我们的发现支持温度和接触模式之间的关系,有助于加深对社会交往与呼吸道传染病流行病学关系的认识。
    BACKGROUND: Human contact patterns are a key determinant driving the spread of respiratory infectious diseases. However, the relationship between contact patterns and seasonality as well as their possible association with the seasonality of respiratory diseases is yet to be clarified.
    METHODS: We investigated the association between temperature and human contact patterns using data collected through a cross-sectional diary-based contact survey in Shanghai, China, between December 24, 2017, and May 30, 2018. We then developed a compartmental model of influenza transmission informed by the derived seasonal trends in the number of contacts and validated it against A(H1N1)pdm09 influenza data collected in Shanghai during the same period.
    RESULTS: We identified a significant inverse relationship between the number of contacts and the seasonal temperature trend defined as a spline interpolation of temperature data (p = 0.003). We estimated an average of 16.4 (95% PrI: 15.1-17.5) contacts per day in December 2017 that increased to an average of 17.6 contacts (95% PrI: 16.5-19.3) in January 2018 and then declined to an average of 10.3 (95% PrI: 9.4-10.8) in May 2018. Estimates of influenza incidence obtained by the compartmental model comply with the observed epidemiological data. The reproduction number was estimated to increase from 1.24 (95% CI: 1.21-1.27) in December to a peak of 1.34 (95% CI: 1.31-1.37) in January. The estimated median infection attack rate at the end of the season was 27.4% (95% CI: 23.7-30.5%).
    CONCLUSIONS: Our findings support a relationship between temperature and contact patterns, which can contribute to deepen the understanding of the relationship between social interactions and the epidemiology of respiratory infectious diseases.
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  • 文章类型: Journal Article
    已知环境暴露与病原体传播和免疫损伤有关,但暴露与病因学和社区获得性肺炎(CAP)严重程度的关系尚不清楚.2014年至2019年在中国8个省的9家医院进行了回顾性观察研究。根据纳入标准招募CAP患者,并采用分子检测方法对呼吸道样本进行33种呼吸道病原体的筛选。社会人口统计学,利用环境和临床因素,结合分布滞后非线性模型,采用logistic回归模型分析病原体检测和疾病严重程度的相关性。共纳入3323例CAP患者,709人(21.3%)患有严重疾病。2064例(62.1%)患者至少一种病原体呈阳性。在阳性组中发现更严重的患者。在调整了混杂因素后,颗粒物(PM)2.5和8-h臭氧(O3-8h)在特定滞后期分别与流感病毒和肺炎克雷伯菌的检测显着相关。PM10和一氧化碳(CO)显示出严重CAP的累积效应。污染物暴露,尤其是PM,O3-8h,在CAP的病原体检测和严重程度中应考虑CO,以提高临床病因和疾病严重程度的诊断。
    Environmental exposures are known to be associated with pathogen transmission and immune impairment, but the association of exposures with aetiology and severity of community-acquired pneumonia (CAP) are unclear. A retrospective observational study was conducted at nine hospitals in eight provinces in China from 2014 to 2019. CAP patients were recruited according to inclusion criteria, and respiratory samples were screened for 33 respiratory pathogens using molecular test methods. Sociodemographic, environmental and clinical factors were used to analyze the association with pathogen detection and disease severity by logistic regression models combined with distributed lag nonlinear models. A total of 3323 CAP patients were included, with 709 (21.3%) having severe illness. 2064 (62.1%) patients were positive for at least one pathogen. More severe patients were found in positive group. After adjusting for confounders, particulate matter (PM) 2.5 and 8-h ozone (O3-8h) were significant association at specific lag periods with detection of influenza viruses and Klebsiella pneumoniae respectively. PM10 and carbon monoxide (CO) showed cumulative effect with severe CAP. Pollutants exposures, especially PM, O3-8h, and CO should be considered in pathogen detection and severity of CAP to improve the clinical aetiological and disease severity diagnosis.
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  • 文章类型: Journal Article
    呼吸道感染对全球公共卫生构成严重威胁,强调迫切需要快速,准确,和大规模的诊断工具。近年来,CRISPR/Cas(成簇的规则间隔短回文重复/CRISPR相关)系统,结合等温扩增方法,在核酸检测(NAT)中得到了广泛的应用。然而,由于重组酶聚合酶扩增(RPA)和CRISPR/Cas试剂之间的竞争作用,实现包含所有必需组分的单管反应系统具有挑战性。此外,为了实现精准医学,区分细菌和病毒感染至关重要。这里,我们开发了一种新的NAT方法,称为一罐RPA-CRISPR/Cas12a,结合了RPA和CRISPR分子诊断技术,能够同时检测12种常见呼吸道病原体,包括六种细菌和六种病毒。RPA和CRISPR/Cas12a反应用石蜡分离,提供用于RPA反应的独立平台以在与CRISPR/Cas12a系统混合之前产生足够的目标产物。结果可以在LED蓝光下目视观察。一锅RPA-CRISPR/Cas12a方法的灵敏度为2.5×100拷贝/μL质粒,与其他细菌或病毒没有交叉反应。此外,通过测试细菌和病毒咽拭子样本的临床分离株来评估临床实用性,表现良好。因此,我们的一锅-RPA-CRISPR/Cas12a方法显示出在即时检测中准确和大规模检测12种常见呼吸道病原体的巨大潜力.
    Respiratory infections pose a serious threat to global public health, underscoring the urgent need for rapid, accurate, and large-scale diagnostic tools. In recent years, the CRISPR/Cas (clustered regularly interspaced short palindromic repeats/CRISPR-associated) system, combined with isothermal amplification methods, has seen widespread application in nucleic acid testing (NAT). However, achieving a single-tube reaction system containing all necessary components is challenging due to the competitive effects between recombinase polymerase amplification (RPA) and CRISPR/Cas reagents. Furthermore, to enable precision medicine, distinguishing between bacterial and viral infections is essential. Here, we have developed a novel NAT method, termed one-pot-RPA-CRISPR/Cas12a, which combines RPA with CRISPR molecular diagnostic technology, enabling simultaneous detection of 12 common respiratory pathogens, including six bacteria and six viruses. RPA and CRISPR/Cas12a reactions are separated by paraffin, providing an independent platform for RPA reactions to generate sufficient target products before being mixed with the CRISPR/Cas12a system. Results can be visually observed under LED blue light. The sensitivity of the one-pot-RPA-CRISPR/Cas12a method is 2.5 × 100 copies/μL plasmids, with no cross-reaction with other bacteria or viruses. Additionally, the clinical utility was evaluated by testing clinical isolates of bacteria and virus throat swab samples, demonstrating favorable performance. Thus, our one-pot-RPA-CRISPR/Cas12a method shows immense potential for accurate and large-scale detection of 12 common respiratory pathogens in point-of-care testing.
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