Multiplex PCR

多重 PCR
  • 文章类型: Meta-Analysis
    Introduction.FilmArray血液培养鉴定面板(BCID)是一种具有高灵敏度和特异性的多重PCR测定法,可鉴定血流感染(BSI)中最常见的病原体。假说。我们假设BCID面板对BSI具有良好的诊断性能,可以在临床应用中推广。目的:为BCID小组用于鉴定阳性血培养的诊断准确性提供总结证据。方法论。我们搜索了MEDLINE,截至2021年3月的EMBASE和Cochrane数据库,并评估了BCID面板诊断测试的有效性。我们进行了荟萃分析,并计算了BCID组的总体敏感性和特异性。系统审查方案已在国际前瞻性系统审查登记册(PROSPERO)(注册号CRD42021239176)中注册。结果。共有16篇全文有资格进行分析。BCID面板对革兰氏阳性细菌的总体敏感性,革兰氏阴性菌和真菌为97%(95%CI,0.96-0.98),100%(95%CI,0.98-01.00)和99%(95%CI,0.87-1.00),分别。合并的诊断特异性为99%(95%CI,0.97-1.00),革兰氏阳性菌100%(95%CI,1.00-1.00)和100%(95%CI,1.00-1.00),革兰氏阴性细菌和真菌,分别。Conclusions.BCID面板对于BSI患者的早期检测具有很高的规则价值。BCID小组仍然可以提供有价值的信息,以排除预测试概率低的人群中的菌血症或真菌血症。
    Introduction. The FilmArray blood culture identification panel (BCID) panel is a multiplex PCR assay with high sensitivity and specificity to identify the most common pathogens in bloodstream infections (BSIs).Hypothesis. We hypothesize that the BCID panel has good diagnostic performance for BSIs and can be popularized in clinical application.Aim: To provide summarized evidence for the diagnostic accuracy of the BCID panel for the identification of positive blood cultures.Methodology. We searched the MEDLINE, EMBASE and Cochrane databases through March 2021 and assessed the efficacy of the diagnostic test of the BCID panel. We performed a meta-analysis and calculated the summary sensitivity and specificity of the BCID panel. Systematic review protocols were registered in the International Prospective Register of Systematic Reviews (PROSPERO) (registration number CRD42021239176).Results. A total of 16 full-text articles were eligible for analysis. The overall sensitivities of the BCID panel on Gram-positive bacteria, Gram-negative bacteria and fungi were 97 % (95 % CI, 0.96-0.98), 100 % (95 % CI, 0.98-01.00) and 99 % (95 % CI, 0.87-1.00), respectively. The pooled diagnostic specificities were 99 % (95 % CI, 0.97-1.00), 100 % (95 % CI, 1.00-1.00) and 100 % (95 % CI, 1.00-1.00) for Gram-positive bacteria, Gram-negative bacteria and fungi, respectively.Conclusions. The BCID panel has high rule-in value for the early detection of BSI patients. The BCID panel can still provide valuable information for ruling out bacteremia or fungemia in populations with low pretest probability.
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  • 文章类型: Meta-Analysis
    背景:抗生素治疗的准确性和时机仍然是下呼吸道感染的挑战。使用多重聚合酶链反应的新分子技术,包括FilmArray®肺炎加面板[FAPP],是为了解决这个问题而开发的。这项研究的目的是在系统评价的荟萃分析中评估FAPP诊断性能,以检测呼吸道样本中的15种典型细菌。
    方法:我们从2010年1月1日至2022年12月31日搜索了PubMed和EMBASE,并选择了与参考标准相比对呼吸样本的FAPP诊断性能的任何研究,细菌培养。主要结果是总体诊断准确性,敏感性和特异性。我们计算了对数诊断赔率比,并分析了单独细菌的性能,抗菌素抗性基因,并根据样品类型。我们还报告了FAPP的周转时间以及面板外细菌的数量和种类。本研究在PROSPERO(CRD42021226280)注册。
    结果:从10317条记录中,我们确定了30项研究,包括8968个样本.21人与重症监护有关。总体敏感性和特异性为94%[95%置信区间(CI)91-95]和98%[95CI97-98],分别。对数诊断赔率比为6.35[95CI6.05-6.65]。在培养物中检测到的9.3%[95CI9.2-9.5]的细菌不包括在FAPP面板中。
    结论:报告FAPP评估的系统评价显示了很高的准确性。该测试可能是肺部细菌感染诊断和抗菌药物管理的辅助工具。需要进一步的证据来评估对临床结果的影响。
    BACKGROUND: Accuracy and timing of antibiotic therapy remain a challenge for lower respiratory tract infections. New molecular techniques using Multiplex Polymerase Chain Reaction, including the FilmArray® Pneumonia Plus Panel [FAPP], have been developed to address this. The aim of this study is to evaluate the FAPP diagnostic performance for the detection of the 15 typical bacteria of the panel from respiratory samples in a meta-analysis from a systematic review.
    METHODS: We searched PubMed and EMBASE from January 1, 2010, to December 31, 2022, and selected any study on the FAPP diagnostic performance on respiratory samples compared to the reference standard, bacterial culture. The main outcome was the overall diagnostic accuracy with sensitivity and specificity. We calculated the log Diagnostic Odds Ratio and analyzed performance for separate bacteria, antimicrobial resistance genes, and according to the sample type. We also reported the FAPP turnaround time and the out-of-panel bacteria number and species. This study is registered with PROSPERO (CRD42021226280).
    RESULTS: From 10 317 records, we identified 30 studies including 8 968 samples. Twenty-one were related to intensive care. The overall sensitivity and specificity were 94% [95% Confidence Interval (CI) 91-95] and 98% [95%CI 97-98], respectively. The log Diagnostic Odds Ratio was 6.35 [95%CI 6.05-6.65]. 9.3% [95%CI 9.2-9.5] of bacteria detected in culture were not included in the FAPP panel.
    CONCLUSIONS: This systematic review reporting the FAPP evaluation revealed a high accuracy. This test may represent an adjunct tool for pulmonary bacterial infection diagnostic and antimicrobial stewardship. Further evidence is needed to assess the impact on clinical outcome.
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  • 文章类型: Journal Article
    在各种情况下,SARS-CoV-2住院患者可发生细菌感染,导致较差的结果,比如更高的死亡率。这项系统评价是为了评估多重PCR检测住院COVID-19患者细菌感染的效率。以及分析最常见的细菌病原体和其他干扰这种诊断的因素。这项研究是使用四个电子数据库(PubMed,泰勒和弗朗西斯,WebofScience,和Wiley在线图书馆)。在290项研究中,9项纳入了系统审查。结果支持使用多重PCR检测细菌,考虑到其高灵敏度和特异性。最常见的细菌病原体是肺炎克雷伯菌,金黄色葡萄球菌,铜绿假单胞菌,肺炎链球菌,和流感嗜血杆菌.入院时的平均年龄为61.5岁,大多数患者是男性(70.3%),在总共1553名患者中。入住ICU的比例很高,在分析的研究中,合并比例为52.6%,平均住院时间为13天。死亡率成比例地高,ICU入院率也是如此,合并死亡率为24.9%。发现65.2%的患者在入院前使用抗生素,有或没有医疗处方。应该有意识地考虑抗生素治疗,考虑到抗生素耐药性的高风险。
    Bacterial infection can occur in patients hospitalized with SARS-CoV-2 in various conditions, resulting in poorer outcomes, such as a higher death rate. This current systematic review was conducted in order to assess the efficiency of multiplex PCR in detecting bacterial infections in hospitalized COVID-19 patients, as well as to analyze the most common bacterial pathogens and other factors that interfere with this diagnosis. The research was conducted using four electronic databases (PubMed, Taylor&Francis, Web of Science, and Wiley Online Library). Out of 290 studies, nine were included in the systematic review. The results supported the use of multiplex PCR in detecting bacteria, considering its high sensitivity and specificity rates. The most common bacterial pathogens found were Klebsiella pneumoniae, Staphylococcus aureus, Pseudomonas aeruginosa, Streptococcus pneumoniae, and Haemophilus influenzae. The median age at admission was 61.5 years, and the majority of patients were men (70.3%), out of a total of 1553 patients. The proportion of ICU admission was very high, with a pooled proportion of 52.6% over the analyzed studies, and an average duration of hospitalization of 13 days. The mortality rate was proportionally high, as was the rate of ICU admission, with a pooled mortality of 24.9%. It was discovered that 65.2% of all patients used antibiotics before admission, with or without medical prescription. Antibiotic treatment should be considered consciously, considering the high risks of developing antibiotic resistance.
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  • 文章类型: Systematic Review
    目的:对于呼吸道病毒的快速样本-反应多重聚合酶链反应(PCR)检测的临床影响尚不明确。我们进行了系统的文献回顾和荟萃分析,以评估在医院环境中可能患有急性呼吸道感染的患者的这种影响。
    方法:我们搜索了EMBASE,MEDLINE,以及2012年至今的Cochrane数据库和2021年的会议程序,用于比较多重PCR测试和标准测试之间的临床影响结果。
    结果:本综述纳入了27项研究,涉及17,321例患者。快速多重PCR检测与-24.22小时(95%CI-28.70至-19.74小时)的结果时间减少相关。住院时间减少-0.82天(95%CI-1.52至-0.11天)。在流感阳性患者中,使用抗病毒药物的可能性更大(RR1.25,95%CI1.06~1.48),在快速多重PCR检测中,使用适当的感染控制设施更为常见(RR1.55,95%CI1.16~2.07).
    结论:我们的系统评价和荟萃分析显示,在流感阳性患者中,患者的总体时间和住院时间以及适当的抗病毒和感染控制管理的改善。该证据支持在医院环境中对呼吸道病毒进行快速样品到答案多重PCR测试的常规使用。
    The clinical impact of rapid sample-to-answer \"syndromic\" multiplex polymerase chain reaction (PCR) testing for respiratory viruses is not clearly established. We performed a systematic literature review and meta-analysis to evaluate this impact for patients with possible acute respiratory tract infection in the hospital setting.
    We searched EMBASE, MEDLINE, and Cochrane databases from 2012 to present and conference proceedings from 2021 for studies comparing clinical impact outcomes between multiplex PCR testing and standard testing.
    Twenty-seven studies with 17,321 patient encounters were included in this review. Rapid multiplex PCR testing was associated with a reduction of - 24.22 h (95% CI -28.70 to -19.74 h) in the time to results. Hospital length of stay was decreased by -0.82 days (95% CI -1.52 to -0.11 days). Among influenza positive patients, antivirals were more likely to be given (RR 1.25, 95% CI 1.06-1.48) and appropriate infection control facility use was more common with rapid multiplex PCR testing (RR 1.55, 95% CI 1.16-2.07).
    Our systematic review and meta-analysis demonstrates a reduction in time to results and length of stay for patients overall along with improvements in appropriate antiviral and infection control management among influenza-positive patients. This evidence supports the routine use of rapid sample-to-answer multiplex PCR testing for respiratory viruses in the hospital setting.
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  • 文章类型: Journal Article
    背景:FilmArray脑膜炎/脑炎(FA/ME)小组在临床实践中带来了好处,但其诊断测试准确性(DTA)仍不清楚。我们旨在确定FA/ME的DTA,用于疑似中枢神经系统(CNS)感染患者的病因诊断。
    方法:我们使用DTA荟萃分析进行了系统评价(PROSPERO:CRD42020139285)。我们搜查了Embase,Medline(Ovid),和WebofScience从成立到9月1日,2021年。我们使用QUADAS-2工具评估了研究级别的偏倚风险,并应用GRADE方法评估了综合证据的确定性。我们包括同时测量参考测试的研究(CSF/血液培养细菌,以及病毒的特异性聚合酶链反应)和疑似CNS感染患者的FA/ME。我们使用CSF/血液培养物(参考测试1)和基于临床/实验室标准的最终诊断裁定(参考测试2)进行了组合敏感性和特异性的随机效应双变量荟萃分析模型。
    结果:我们纳入了19项研究(11,351名参与者)。对于具有参考测试1(16项研究/6183名患者)的所有细菌,敏感性估计为89·5%(95CI81·1-94·4),特异性为97·4%(95CI94-98·9)。参考测试2(15项研究/5,524名患者),对于单纯疱疹病毒2(HSV-2),敏感性估计为92·1%(95CI86·8-95·3),特异性为99.2(95CI98·3-99·6),肠病毒,水痘-带状疱疹病毒(VZV),我们获得的灵敏度在75·5和93·8%之间,和超过99%的特异性(参考测试1)。证据的确定性很低。
    结论:FA/ME可能对鉴定细菌具有可接受的至高的敏感性和高特异性,尤其是肺炎链球菌,和病毒,特别是对于HSV-2和肠道病毒。对单核细胞增生性弧菌的敏感性,流感嗜血杆菌,大肠杆菌,HSV-1次优。
    背景:无。
    BACKGROUND: The FilmArray Meningitis/Encephalitis(FA/ME) panel brings benefits in clinical practice, but its diagnostic test accuracy (DTA) remains unclear. We aimed to determine the DTA of FA/ME for the aetiological diagnostic in patients with suspected central nervous system(CNS) infection.
    METHODS: We performed a systematic review with DTA meta-analysis (PROSPERO: CRD42020139285). We searched Embase, Medline (Ovid), and Web of Science from inception until September 1st, 2021. We assessed the study-level risk of bias with the QUADAS-2 tool and applied the GRADE approach to assess the certainty of the synthesised evidence. We included studies that simultaneously measured the reference test (CSF/blood culture for bacteria, and specific polymerase chain reaction for viruses) and the FA/ME in patients with suspected CNS infection. We performed random-effects bivariate meta-analysis models of combined sensitivity and specificity using CSF/blood cultures(reference test 1) and a final diagnosis adjudication based on clinical/laboratory criteria (reference test 2).
    RESULTS: We included 19 studies (11,351 participants). For all bacteria with reference test 1 (16 studies/6183 patients) sensitivity was estimated at 89·5% (95%CI 81·1-94·4), and specificity at 97·4% (95%CI 94-98·9). With reference test 2 (15 studies/5,524 patients), sensitivity was estimated at 92·1%(95%CI 86·8-95·3) and specificity at 99.2(95%CI 98·3-99·6) For herpes simplex virus-2(HSV-2), enteroviruses, and Varicella-Zoster virus (VZV), we obtained sensitivities between 75·5 and 93·8%, and specificities above 99% (reference test 1). Certainty of the evidence was low.
    CONCLUSIONS: FA/ME may have acceptable-to-high sensitivities and high specificities for identifying bacteria, especially for S.pneumoniae, and viruses, especially for HSV-2, and enteroviruses. Sensitivities for L.monocytogenes, H.influenzae, E.coli, and HSV-1 were suboptimal.
    BACKGROUND: None.
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  • 文章类型: Journal Article
    哺乳动物物种鉴定是法医学的重要问题之一。确定在犯罪现场发现的非人类生物材料的来源可以通过缩小嫌疑人的范围来增加识别真正罪魁祸首的可能性。尽管已经开发了许多基于线粒体DNA(mtDNA)的技术,从含有来自多个物种的DNA混合物的降解样品中经济有效地鉴定物种以及标准化哺乳动物物种鉴定程序仍然存在挑战。这篇综述评估了基于mtDNA的技术的可靠性和多功能性,以揭示建立标准分析方法的障碍,特别关注DNA混合物。当样品含有来自多个物种的DNA混合物时,测序分析结果的解释是困难的。尽管使用下一代测序(NGS)技术的DNA元编码可以克服DNA混合问题,DNA元编码不适合当地法医实验室常规进行的小规模分析类型,主要是因为它既昂贵又耗时。相比之下,荧光多重PCR分析能够从次优样品中进行经济有效的同时物种鉴定,尽管与测序技术相比,目前可识别物种的数量有限。本综述中介绍的当前技术的优点和局限性表明,多重PCR分析对于法医案例分析中的哺乳动物物种鉴定将继续很重要。多重PCR分析的进一步发展,能够鉴定更多的物种,将成为法医实验室标准化工作的关键步骤。
    Mammalian species identification is one of the important issues in forensic science. Determining the origins of non-human biological material found at crime scenes can increase the possibility of identifying the true culprit by narrowing down the range of suspects. Although many techniques based on mitochondrial DNA (mtDNA) have been developed, challenges remain to cost-effectively identify species from degraded samples containing a mixture of DNA from multiple species and to standardize procedures for mammalian species identification. This review evaluates the reliability and versatility of mtDNA-based techniques to reveal obstacles to the establishment of standard analytical methods, with a particular focus on DNA mixtures. When samples contain a mixture of DNA from multiple species, the interpretation of sequencing analysis results is difficult. Although DNA metabarcoding using next-generation sequencing (NGS) technologies can overcome the DNA mixture problem, DNA metabarcoding is not suitable for the type of small-scale analysis routinely performed by local forensic laboratories, primarily because it is costly and time-consuming. By contrast, fluorescent multiplex PCR analysis enables cost-effective and simultaneous species identification from suboptimal samples, although the number of identifiable species is currently limited in comparison with sequencing techniques. The advantages and limitations of current techniques presented in this review indicate that multiplex PCR analysis will continue to be important for mammalian species identification in forensic casework analysis. Further developments in multiplex PCR analysis that enable the identification of an increased number of species will play a key step for standardization efforts among forensic laboratories.
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  • 文章类型: Journal Article
    BACKGROUND: The FilmArray® meningitis/encephalitis (ME) panel is a multiplex PCR assay which can detect the most commonly identified pathogens in central nervous system infections. It significantly decreases the time to diagnosis of ME and data has yielded several positive outcomes. However, in part, reports of both false positive and false negative detections have resulted in concerns about adoption.
    OBJECTIVE: The aim was to evaluate the ME panel in a diagnostic test accuracy review.
    METHODS: The PubMed and EMBASE databases were systematically searched through May 2019.
    METHODS: Eligible studies were those providing sensitivity and specificity data for the ME panel compared with a reference standard. Studies providing details on false positive and false negative results of the panel as well as further investigation (adjudication) of the discordant results between the panel and comparator assays were included and assessed separately.
    METHODS: Patients with suspected ME for whom a panel was ordered were included.
    METHODS: The ME panel was compared to reference standard methods for diagnosing community-acquired ME. We performed a meta-analysis and calculated the summary sensitivity and specificity of the ME panel. Moreover, we evaluated the false positive and false negative results of the panel.
    RESULTS: Thirteen studies (3764 patients) were included in the review and 8 of them (3059 patients) were pooled in a meta-analysis. The summary estimates of sensitivity and specificity with 95% confidence intervals (CI) was 90% (95% CI 86-93%) and 97% (95% CI 94-99%), respectively. When we looked specifically at studies that assessed further the false positive and false negative results, false positive detections were 11.4% and 4% before and after adjudication, respectively. The highest proportion of false positive was observed for Streptococcus pneumoniae followed by Streptococcus agalactiae. False negative isolates were 2.2% and 1.5% before and after adjudication, respectively. Herpes simplex virus 1 and 2, enterovirus and Cryptococcus neoformans/gattii had the highest proportions of false negative determinations. False negative C. neoformans/gattii were mostly patients with positive antigen titres, on treatment or cleared disease.
    CONCLUSIONS: The currently available literature suggests that the ME panel has high diagnostic accuracy. However, the decision for implementation should be individualized based on the needs of the patient population, the capabilities of the laboratory, and the knowledge of the healthcare providers that will utilize the test.
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  • 文章类型: Journal Article
    尽管Cyberlindnerafabinaii是一种罕见的机会主义酵母,导致败血症的能力,产生生物膜,快速获得对氟康唑和伏立康唑的耐药性,加强了从其密切相关的物种中识别它的冲动。广泛使用的生化测定法主要将Cyberlindnerafabinaii鉴定为Cyberlindnerajadinii和Wickerhamomyces,导致在临床上低估了这种酵母。此外,28年来,对可靠分子鉴定手段的渴望仍未解决。为了明确区分Cy。Fabianii,Cy.密西西比州,Cy.Jadinii,和W.异常,我们设计了一种双功能多重聚合酶链反应(PCR)检测方法。挑战我们的双功能多重PCR检测与30个临床上最重要的酵母物种,证明了它的特殊性。尽管常规PCR可以区分四种目标物种,由于Tm重叠导致的实时PCR对应物错误鉴定了Cy。密西西比州作为Cy。Jadinii.除了对已发表的Cy案例进行全面的文献综述。从1990年到2018年,我们从德黑兰收集了各种临床分离株,设拉子,和Fasa(2017年7月1日至2017年12月31日),以发现这些密切相关的物种在伊朗的被动相对分布。对我们的伊朗酵母分离株进行基质辅助激光解吸/电离飞行时间(MALDI-TOF)MS和LSU以及ITSrDNA测序,发现了六个Cy分离株。fabianii(中心静脉导管n=2,阴道拭子n=4)和一个分离的Cy。jadinii(阴道拭子)。由于在全球ARTEMIS研究中使用生化测定,我们鼓励重新鉴定Cy的临床分离株。Jadinii和Cy.jadinii使用MALDI-TOF或Sanger测序可能导致纠正这种真菌的分布。
    Although Cyberlindnera fabinaii is a rare opportunist yeast species, its ability to cause septicemia, produce biofilm, and rapid acquisition of resistance to fluconazole and voriconazole, reinforced the urge for its identification from its closely related species. Widely used biochemical assays mainly identify Cyberlindnera fabinaii as Cyberlindnera jadinii and Wickerhamomyces anomalus, resulting in underestimation of this yeast in clinical settings. Moreover, the urge for a reliable molecular means of identification remains unsolved for 28 years. In order to unequivocally differentiate Cy. fabianii, Cy. mississipiensis, Cy. jadinii, and W. anomalus, we designed a dual-function multiplex polymerase chain reaction (PCR) assay. Challenging our dual-function multiplex PCR assay with 30 most clinically important yeast species, proved its specificity. Although conventional PCR could differentiate four target species, the real-time PCR counterpart due to Tm overlap misidentified Cy. mississipiensis as Cy. jadinii. Alongside of presenting a comprehensive literature review of published cases of Cy. fabianii from 1990 to 2018, we collected various clinical isolates from Tehran, Shiraz, and Fasa (July 1, 2017, to December 31, 2017) to find a passive relative distribution of these closely-related species in Iran. Subjecting our Iranian collection of yeast isolates to matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) MS and LSU and ITS rDNA sequencing revealed six isolates of Cy. fabianii (central venous catheter n = 2 and vaginal swabs n = 4) and one isolate of Cy. jadinii (vaginal swabs). Due to the use of biochemical assays in global ARTEMIS study, we encourage reidentification of clinical isolates of Cy. jadinii and Cy. jadinii using MALDI-TOF or Sanger sequencing that might lead to correcting the distribution of this fungus.
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  • 文章类型: Journal Article
    OBJECTIVE: To provide a summary of evidence for the diagnostic accuracies of three multiplex PCR systems (mPCRs)-BioFire FilmArray RP (FilmArray), Nanosphere Verigene RV+ test (Verigene RV+) and Hologic Gen-Probe Prodesse assays-on the detection of viral respiratory infections.
    METHODS: A comprehensive search up to 1 July 2017 was conducted on Medline and Embase for studies that utilized FilmArray, Verigene RV+ and Prodesse for diagnosis of viral respiratory infections. A summary of diagnostic accuracies for the following five viruses were calculated: influenza A virus (FluA), influenza B virus, respiratory syncytial virus, human metapneumovirus and adenovirus. Hierarchical summary receiver operating curves were used for estimating the viral detection performance per assay.
    RESULTS: Twenty studies of 5510 patient samples were eligible for analysis. Multiplex PCRs demonstrated high diagnostic accuracy, with area under the receiver operating characteristic curve (AUROC) equal to or more than 0.98 for all the above viruses except for adenovirus (AUROC 0.89). FilmArray, Verigene RV+ and ProFlu+ (the only Prodesse assay with enough data) demonstrated a summary sensitivity for FluA of 0.911 (95% confidence interval, 0.848-0.949), 0.949 (95% confidence interval, 0.882-0.979) and 0.954 (95% confidence interval, 0.871-0.985), respectively. The three mPCRs were comparable in terms of detection of FluA.
    CONCLUSIONS: Point estimates calculated from eligible studies showed that the three mPCRs (FilmArray, Verigene RV+ and ProFlu+) are highly accurate and may provide important diagnostic information for early identification of respiratory virus infections. In patients with low pretest probability for FluA, these three mPCRs can predict a low possibility of infection and may justify withholding empirical antiviral treatments.
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