Rapid determination of Proteus mirabilis susceptibility to antibiotics using infrared spectroscopy in tandem with random forest

Bacterial infections cause serious illnesses that are treated with antibiotics. Currently used methods for detecting bacterial antibiotic susceptibility consume 48–72 h, leading to overuse of antibiotics. Thus, many bacterial species have acquired resistance to a broad range of available antibiotics. There is an urgent need to develop efficient methods for rapid determination of bacterial susceptibility to antibiotics. The combination of machine learning and Fourier-transform infrared (FTIR) spectroscopy has generated a promising diagnostic approach in medicine and biology. Our main goal is to examine the potential of FTIR spectroscopy to determine the susceptibility of urinary tract infection-Proteus mirabilis to a specific range of antibiotics, within about 20 min after 24 h culture and identification. We measured the infrared spectra of 489 different P. mirabilis isolates and used random forest to analyze this spectral database. A classification success rate of ~84% was achieved in differentiating between the resistant and sensitive isolates based on their susceptibility to ceftazidime, ceftriaxone, cefuroxime, cefuroxime axetil, cephalexin, ciprofloxacin, gentamicin, and sulfamethoxazole antibiotics in a time span of 24 h instead of 48 h.     

A novel diagnostic method for human immunodeficiency virus type-1 in plasma by near-infrared spectroscopy

Presently, the diagnosis of virus infections is based mainly on serological assays. Although polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA) have been increasingly used for the diagnosis of such viral infections, the risk of transfusion-transmitted blood-borne viruses remains. Furthermore, PCR and ELISA are expensive and time-consuming, and sometimes cause falsepositive or false-negative results. Therefore, a rapid, accurate and cost-effective diagnostic procedure is needed. We subjected plasma from individuals infected with human immunodeficiency virus type-1 (HIV-1), the causative agent of acquired immune deficiency syndrome (AIDS), as well as plasma from uninfected individuals as a control to near-infrared (NIR) spectroscopy, which may provide a rapid diagnostic method for HIV-1 infection without using any reagent. NIR spectra in the 600-1,000 nm region for plasma from pre-serologically HIV-1-infected individuals and healthy donors were subjected to partial least squares (PLS) regression analysis and leave-out cross-validation to develop a multivariate model to estimate the concentration of HIV-1. Simultaneously, the same plasma samples were examined for HIV-1 p24 by ELISA. The results obtained by the NIR spectroscopy model for HIV-1 yielded a good correlation with those obtained by the reference method (HIV-1 p24 ELISA). These results suggest that NIR spectroscopy using plasma could provide a rapid, accurate, cost-effective tool for large-scale diagnosis of HIV-1 infection.     

病毒性肺炎患者外周血干扰素刺激基因15 mRNA表达特征的研究

通过分析病毒性肺炎患者外周血干扰素刺激基因15(interferon stimulated gene 15,ISG15)mRNA在感染状态下的特征性表达,探究ISG15与病毒性肺炎发病机制及疾病活动性之间的可能联系,为探索特异性呼吸道病毒感染的宿主生物标志物提供依据.本研究共收集157例社区获得性肺炎(communit...     

中性粒细胞CD64指数、PCT及NLR在 感染性疾病诊断中的价值

目的探讨中性粒细胞CD64指数、降钙素原(PCT)和中性粒细胞与淋巴细胞比值(NLR)在感染性疾病诊断和鉴别诊断中的临床价值。方法选取确诊的感染性疾病患者127例和同期健康对照者54例为研究对象,感染性疾病患者按照血培养和微生物检测结果分为病毒感染组(51例)和细菌感染组(76例)。分别检测每一位受试对象的外周血中性粒细胞CD64指数、PCT水平和NLR,应用受试者工作特征曲线(ROC)研究这3项指标在感染性疾病诊断和鉴别中的临床价值。结果感染性疾病患者中性粒细胞CD64指数、PCT水平和NLR明显高于健康对照组(P0.01)。细菌感染组患者中性粒细胞CD64指数、PCT水平和NLR明显高于病毒感染组(P0.01)。ROC曲线显示:当用于诊断感染性疾病时,中性粒细胞CD64指数、PCT和NLR的AUC值分别为0.799、0.655、0.622,灵敏度分别为75.4%、63.5%、60.3%,特异度分别为79.6%、67.4%、60.0%,3项指标联合检测的AUC值为0.812;当用于鉴别诊断细菌感染和病毒感染时,中性粒细胞CD64指数、PCT和NLR的AUC值分别为0.931、0.728、0.551,灵敏度分别为89.1%、70.3%、60.9%,特异度分别为80.0%、64.6%、57.2%,3项指标联合检测的AUC值为0.935。结论在感染性疾病的诊断和鉴别感染类型中,中性粒细胞CD64指数明显优于PCT和NLR。3项指标联合检测优于任何一个单项指标的检测效果。     

Development and validation of multiplex real-time PCR assays for rapid detection of cytomegalovirus,Epstein-Barr virus,and polyomavirus BK in whole blood from transplant candidates

Transplant recipients are more susceptible to bacterial and viral infections. Cytomegalovirus (CMV), Epstein-Barr virus (EBV), and polyomavirus BK (BK) are risk factors for graft dysfunction. All three of them are latent viruses that can cause serious disease in immunocompromised patients. Mainly qualitative PCR tests are required for diagnosis and quantitative monitoring, which are used to follow the response to transplantation. We developed a multiplex real-time PCR (qPCR) method to detect these viruses during blood screenings of transplant recipients. We also validated analytical and clinical performance tests using the developed multiplex qPCR. The limit of detection (LOD) was 100, 125, and 183 copies/ml for CMV, EBV, and BK, respectively. These results had high linearity (R² = 0.997) and reproducibility (CV range, 0.95–2.38%, 0.52–3.32%, and 0.31–2.45%, respectively). Among 183 samples, we detected 8 samples that were positive for CMV, while only 6 were positive for EBV, and 3 were positive for BK. Therefore, the viral infection prevalence in transplant candidates was 4.40% for CMV, 3.29% for EBV, and 1.64% for BK. This multiplex qPCR method should be used widely for diagnosing and monitoring latent viral infections in transplant recipients.     

血清降钙素原在开颅手术后颅内感染诊断及预后中的诊断价值

目的探讨血清降钙素原（PCT）在开颅手术后并发颅内感染患者诊断中的价值及预后评估应用。方法回顾性分析五年来我院神经外科开颅手术后并发颅内感染患者115例,根据颅内感染的病原体学诊断和临床诊断标准分为细菌性感染组45例和非细菌性感染组70例,分别对两组患者进行血清检测PCT,C反应蛋白（CRP）值及预后分析,采用成组T检验比较两组数据的差异。结果开颅手术后并发颅内细菌感染患者血清PCT值显著高于非细菌性感染组,差异有统计学意义（t=15．43,P〈0．01）;而CRP的增高在两组之间差异无统计学意义（t=1．83,P〉0．05）。细菌性颅内感染的PCT浓度主要分布在2＋区域,即2—10ng／mL,而在非细菌性感染中,主要分布在阴性区域,即〈0．5ng／mL。而在细菌感染组的预后上,PCT〉10ng／mL患者死亡率显著高于PCT〈10ng／mL的感染患者（X2=7．2,P〈0．01）。结论监测PCT对开颅手术后并发颅内感染患者具有明显的临床诊断价值。此外,PCT值的高低有助于评估颅内细菌感染的预后。     

On the epidemiology and etiology of pneumonia in adults

In a group of 74 hospitalized patients with the diagnosis of acute infectious pneumonia, the etiological contribution of viral and bacterial agents is analyzed in cases of clarified etiology and an assessment is made of the relationship between the explained etiology and the overall epidemiological situation. Etiology was clarified in 36 patients (48.6%). Viral and bacterial etiology was confirmed in 13.3% and 39.8% of the entire group respectively. In three cases, mixed viral and bacterial infection was reported. Most prominent among the viral agents were herpes simplex, parainfluenza, respiratory syncytial and influenza type B viruses. As far as the bacterial agents were concerned, the species most frequently isolated were Staphylococcus aureus, Streptococcus pneumoniae and a variety of Enterobacteriaceae. The relationship between the overall epidemiological situation and pneumonia etiology is discussed as well as the relevance of the diagnostic methods employed.     

Restrictive antibiotherapy after renal transplantation.

Forty-two patients were followed up after 44 renal transplantations in an effort to evaluate possible benefits from the following protocol: systematic microbiologic and clinical surveillance, early and aggressive research for the cause of suspected infections, refusal to use prophylactic antibiotherapy, and selection of treatment according to the established cause of the infection. During 18,030 days of follow-up 124 infections were recorded, of which 110 were bacterial, 11 viral and 3 protozoal. Eighty originated in the urinary tract, 17 in skin wounds and 10 in the lower respiratory tract. Septicemia occurred three times, and one death due to infection was recorded. In the treatment of bacterial infections patients received antibiotics for 2486 days. Ampicillin (given for 816 days) and "minor" drugs such as sulfonamides and urinary antiseptics (given for 1036 days) were used 74.5% of the time, whereas gentamicin was used only 2.6% of the time (64 days). Combined antibacterial therapy was needed 1.2% of the time (29 days). A restrictive policy regarding anti-biotherapy seems to be beneficial to renal transplant recipients.     

Molecular diagnostic assays for detection of viral respiratory pathogens in institutional outbreaks

Outbreaks of viral respiratory disease in institutions may be associated with high morbidity and mortality, depending upon the viral etiology and the age and immune status of the affected patients. Control of outbreaks may include isolation and/or cohorting, and prophylaxis or treatment with specific antiviral agents may be indicated, all dependent upon the specific cause of the outbreak. Conventional methods of viral diagnosis detect only a limited number of the viruses that are known to cause outbreaks. The availability of sensitive and specific molecular assays has facilitated rapid diagnosis of a wider range of viruses from respiratory outbreaks. Molecular methods have distinct advantages over conventional methods, including the ability to rapidly develop assays for emerging viruses and new variants of existing viruses. In addition, molecular testing allows rapid detection of resistance to antiviral agents or mutations leading to increased virulence. However, high-throughput molecular testing requires batch processes that may compromise the ability to respond quickly to urgent testing demands.     

Cystic fibrosis infections: treatment strategies and prospects

Pseudomonas aeruginosa and Burkholderia cepacia are the two major Gram-negative rods that colonize/infect the lungs of patients with cystic fibrosis (CF). These organisms may cause progressive respiratory failure, although occasionally more rapid infections result in the ' Cepacia ' syndrome. Many antibiotics have been used against Pseudomonas and Burkholderia , but once chronic colonization has been established, eradication of these organisms is rare. Drug therapy for CF patients is compromised by a number of bacterial factors that render the infectious agents resistant to antibiotics, including efflux pumps that remove antibiotics, lack of penetration of antibiotics into bacterial biofilms, and changes in the cell envelope that reduce the permeability of antibiotics. Any combination of these mechanisms increases the likelihood of bacterial survival. Therefore, combinations of antibiotics or of antibiotic and nonantibiotic compounds are currently being tested against Pseudomonas and Burkholderia . However, progress has been slow, with only occasional combinations showing promise for the eradication of persistent Gram-negative rods in the airways of CF patients. This review will summarize the current knowledge of CF infections and speculate on potential future pathways to treat these chronic infections.     

Overview of vaccines and vaccination

Of the 80-plus known infectious agents pathogenic for humans, there are now more than 30 vaccines against 26 mainly viral and bacterial infections and these greatly minimize subsequent disease and prevent death after exposure to those agents. This article describes the nature of the vaccines, from live attenuated agents to subunits, their efficacy and safety, and the kind of the immune responses generated by those vaccines, which are so effective. To date, all licensed vaccines generate especially specific antibodies, which attach to the infectious agent and therefore can very largely prevent infection. These vaccines have been so effective in developed countries in preventing mortality after a subsequent infection that attempts are being made to develop vaccines against many of the remaining infectious agents. Many of the latter are difficult to manipulate; they can cause persisting infections or show great antigenic variation. A range of new approaches to improve selected immune responses, such as immunization with DNA or chimeric live vectors, viral or bacterial, are under intense scrutiny, as well as genomic analysis of the agent.     

In vivo dynamic characterization of the human tympanic membrane using pneumatic optical coherence tomography

Decreased mobility of the human eardrum, the tympanic membrane (TM), is an essential indicator of a prevalent middle ear infection. The current diagnostic method to assess TM mobility is via pneumatic otoscopy, which provides subjective and qualitative information of subtle motion. In this study, a handheld spectral-domain pneumatic optical coherence tomography system was developed to simultaneously measure the displacement of the TM, air pressure inputs applied to a sealed ear canal, and to perform digital pneumatic otoscopy. A novel approach based on quantitative parameters is presented to characterize spatial and temporal variations of the dynamic TM motion. Furthermore, the TM motions of normal middle ears are compared with those of ears with middle ear infections. The capability of noninvasively measuring the rapid motion of the TM is beneficial to understand the complex dynamics of the human TM, and can ultimately lead to improved diagnosis and management of middle ear infections.     

Infectious exacerbations of chronic obstructive pulmonary disease associated with respiratory viruses and non-typeable Haemophilus influenzae

Infectious exacerbations of chronic obstructive pulmonary disease (COPD) have been reported to occur with both viral and bacterial pathogens. In this study, 35 exacerbations associated with the isolation of non-typeable Haemophilus influenzae from sputum were identified as part of a prospective longitudinal study. Samples from these patients were subjected to immunoassays to identify a new immune response to the homologous isolate of non-typeable H. influenzae to more accurately assess a bacterial etiology. These patients also were studied carefully for evidence of viral infection using viral culture, serology and polymerase chain reaction-based assays. Sixteen of 35 exacerbations (45.7%) were associated with evidence of acute viral infection and 11 of the 35 exacerbations (31.4%) were associated with the development of new serum IgG to homologous non-typeable H. influenzae. Overall, evidence of infection with a respiratory virus or non-typeable H. influenzae was seen in 24 of 35 exacerbations (68.6%). No association between viral infection and immune response to non-typeable H. influenzae was observed, although a trend toward an immune response to non-typeable H. influenzae and absence of viral infection was seen. The results show that exacerbations in adults with COPD were associated with infection caused by virus alone, non-typeable H. influenzae alone, or virus and non-typeable H. influenzae simultaneously.     

890例疑似细菌性阴道病患者阴道分泌物检测结果与分析

目的探讨BV三项检测法在阴道感染诊断中的价值。方法对890例阴道分泌物进行常规检查和BV三项检测。结果 890例疑似细菌性阴道病患者中,BV三项检测法检出BV患者772例(占总数的86.74%),常规镜检(Am sel标准法)检出BV患者763例(占总数的85.73%),二者的符合率为93.40%,具有中度一致性(Kappa=0.746,P0.05)。890例疑似细菌性阴道病患者中,BV合并VVC感染患者129例(占总数的14.49%),BV合并TV感染患者41例(占总数的4.61%),混合感染(BV+VVC、BV+TV)的发病率达到了19.10%(170/890)。结论 BV三项检测法不仅可初筛混合感染,而且与Am sel标准法在诊断BV方面具有较高的一致性,该检测方法快速准确,适宜临床推广与应用。     

拉曼光谱技术在病原菌感染诊断中的应用进展

快速准确诊断感染性疾病病原体是遏制超级细菌传播和抗生素滥用的重要防线。目前,临床病原菌感染诊断十分依赖于培养手段,导致检测周期长达数日,不但影响了患者的及时诊治,还间接导致抗生素的滥用。拉曼光谱技术是一种无损、高灵敏的分子指纹图谱检测技术,近年来在生物学领域得到广泛应用,其具有的免培养、快速、高特异性、低成本等优点为病原菌感染的诊断提供了新方案。本文阐述了拉曼光谱技术的原理和特点,综述了其在病原菌鉴定和抗菌药物敏感性试验方面的应用进展。     

The role of respiratory tract viruses in the etiology of obstructive bronchitis in infants

Out of 524 children with acute respiratory infections in 141 obstructive bronchitis was diagnosed (OZO). Seventy cases could be linked to viral infection. Viral infections tested (influenza virus A, B, parainfluenza typ 1-3, RSV, adenoviruses) were more frequently associated with OZO than other acute respiratory infections of unknown etiology. Majority infections induced by influenza virus A and parainfluenza virus typ 2 were accompanied by OZO symptoms. Of the highest risk of acquiring OZO despite of viral infection participation, were children of 4-12 months of age. OZO associated viral infections prevailed during autumn-winter season, while in spring-summer period undetermined factors were the major cause of OZO. In serum samples of children with OZO, despite of etiology of the disease, higher level of IgE was found than in a group of children without the symptoms. In the case of OZO of unestablished etiology the level of serum IgE was significantly higher than in the cases when viral etiology of the disease was found.     

Pathogens in children with severe combined immune deficiency disease or AIDS.

We evaluated the frequency and severity of illnesses caused by various microbial pathogens in 15 children with severe combined immune deficiency disease (SCID) and 8 with acquired immune deficiency syndrome (AIDS). There were 35 viral, 23 bacterial, 19 mycotic and 13 parasitic infections. Nineteen of the 23 patients died of infection; Pneumocystis carinii pneumonia, giant-cell pneumonia due to paramyxoviruses and various disseminated viral infections were responsible for most deaths in both groups. The emerging role of paramyxoviruses was illustrated by the fact that they were responsible for giant-cell pneumonia in seven patients. Viral enteric infections were frequent in both groups. The variety of infectious microorganisms and the severity of resulting illnesses in the patients with AIDS were similar to those in the patients with SCID.     

Influence of Dendritic Cells on Viral Pathogenicity

Although most viral infections cause minor, if any, symptoms, a certain number result in serious illness. Viral disease symptoms result both from direct viral replication within host cells and from indirect immunopathological consequences. Dendritic cells (DCs) are key determinants of viral disease outcome; they activate immune responses during viral infection and direct T cells toward distinct T helper type responses. Certain viruses are able to skew cytokine secretion by DCs inducing and/or downregulating the immune system with the aim of facilitating and prolonging release of progeny. Thus, the interaction of DCs with viruses most often results in the absence of disease or complete recovery when natural functions of DCs prevail, but may lead to chronic illness or death when these functions are outmanoeuvred by viruses in the exploitation of DCs.     

An infectious aetiology of sudden infant death syndrome

Due attention has been given to infectious agents and immune responses to infection in sudden infant death syndrome (SIDS). It has been acknowledged that the pathological, epidemiological and genotypic findings in SIDS infants suggest an infectious aetiology possibly being potentiated by immunoregulatory polymorphisms, however, the cause of SIDS is a mystery and remains open to debate. Consistent pathological findings are seen which display similarities to the pathogenesis of toxaemic shock and/or sepsis. The major risk factors for SIDS parallel those for increased colonization and serious bacterial infections and the natural variation in the incidence of SIDS cases is typical of an infectious disease. The roles played by viral infection, immunoregulatory genes and suspected bacterial species are discussed herein.