A technological update of molecular diagnostics for infectious diseases

Identification of a causative pathogen is essential for the choice of treatment for most infectious diseases. Many FDA approved molecular assays; usually more sensitive and specific compared to traditional tests, have been developed in the last decade. A new trend of high throughput and multiplexing assays are emerging thanks to technological developments for the human genome sequencing project. The applications of microarray and ultra high throughput sequencing technologies for diagnostic microbiology are reviewed. The race for the $1000 genome technology by 2014 will have a profound impact in diagnosis and treatment of infectious diseases in the near future.     

Antibodies as delivery vehicles for radioimmunotherapy of infectious diseases

The field of infectious diseases is in crisis and there is a need for strategies that can facilitate the rapid development of new antimicrobial agents. Radioimmunotherapy (RIT), a therapeutic modality originally developed for cancer treatment, has recently been suggested as a novel therapy for the treatment of a variety of infectious diseases. Because specific antibodies are used in RIT as delivery vehicles of cytocidal radiation, their molecular weight influences the nonspecific accumulation in infectious foci and blood clearance, and their affinity-specific accumulation of antibodies in infectious foci. Like the problems encountered in oncology, relevant variables in the development of RIT of infectious diseases include target antigen-shedding; delivering radionuclides to infectious foci in organs, abscesses, granulomas, heart and brain, and potential safety concerns. Dadachova and Casadevall anticipate that RIT can be developed for many types of infectious diseases, including microbes resistant to conventional antimicrobial therapy and agents of biological warfare.     

Antibodies as delivery vehicles for radioimmunotherapy of infectious diseases

The field of infectious diseases is in crisis and there is a need for strategies that can facilitate the rapid development of new antimicrobial agents. Radioimmunotherapy (RIT), a therapeutic modality originally developed for cancer treatment, has recently been suggested as a novel therapy for the treatment of a variety of infectious diseases. Because specific antibodies are used in RIT as delivery vehicles of cytocidal radiation, their molecular weight influences the nonspecific accumulation in infectious foci and blood clearance, and their affinity-specific accumulation of antibodies in infectious foci. Like the problems encountered in oncology, relevant variables in the development of RIT of infectious diseases include target antigen-shedding; delivering radionuclides to infectious foci in organs, abscesses, granulomas, heart and brain, and potential safety concerns. Dadachova and Casadevall anticipate that RIT can be developed for many types of infectious diseases, including microbes resistant to conventional antimicrobial therapy and agents of biological warfare.     

Recent advances in the administration of vaccines for infectious diseases: microneedles as painless delivery devices for mass vaccination

Despite remarkable progress in the control of infectious diseases through vaccination, better delivery systems have been poorly explored. There is renewed interest in the discovery of novel vaccines and adjuvants owing to emerging and reemerging diseases and the burden and complexity of chronic infectious diseases. Conversely, the need for rapid local, regional, mucosal or parenteral bioavailability has led to advances in delivery systems and devices. Here, we present recent developments in the field of non-invasive cutaneous delivery of vaccines for infectious diseases. Transdermal delivery using microneedles could revolutionize the way prophylactic interventions for infectious diseases are carried out in the future.     

Rapid analysis of pharmacology for infectious diseases

Pandemic, epidemic and endemic infectious diseases are united by a common problem: how do we rapidly and cost-effectively identify potential pharmacological interventions to treat infections? Given the large number of emerging and neglected infectious diseases and the fact that they disproportionately afflict the poorest members of the global society, new ways of thinking are required to developed high productivity discovery systems that can be applied to a larger number of pathogens. The growing availability of parasite genome data provides the basis for developing methods to prioritize, a priori, the potential drug target and pharmacological landscape of an infectious disease. Thus the overall objective of infectious disease informatics is to enable the rapid generation of plausible, novel medical hypotheses of testable pharmacological experiments, by uncovering undiscovered relationships in the wealth of biomedical literature and databases that were collected for other purposes. In particular our goal is to identify potential drug targets present in a pathogen genome and prioritize which pharmacological experiments are most likely to discover drug-like lead compounds rapidly against a pathogen (i.e. which specific compounds and drug targets should be screened, in which assays and where they can be sourced). An integral part of the challenge is the development and integration of methods to predict druggability, essentiality, synthetic lethality and polypharmacology in pathogen genomes, while simultaneously integrating the inevitable issues of chemical tractability and the potential for acquired drug resistance from the start.     

Evolving importance of biologics and novel delivery systems in the face of microbial resistance

Methods to control infectious diseases in livestock are growing in importance. As the size of the average farm increases-for poultry, dairy and beef cattle, swine, and fish-the risk of rapid spread of infectious diseases increases as well. This increases the need for alternative methods of control of infectious agents. Improvements in specific immunogens, adjuvants, and delivery systems are needed to meet the demand for vaccines to ensure a healthy and safe meat supply. This article explores the challenges, trends, and recent advances in the control of infectious diseases through the use of biologics.     

Electrochemiluminescence in bioanalysis

The discovery of electrochemiluminescence (ECL) and its development as a means of detection is truly a success story. Although studies describing ECL were published in the early 1960s, most studies using ECL as a means of detection were not widely published until the mid 1990s. Incorporating ECL into assays provides increased sensitivity, several logs of dynamic range and the ability to electronically control the reaction. These characteristics provide advantages over assays that rely on radioisotopic labels, fluorescence and enzymatic activity. There have been many areas of science that have benefited from the use of ECL, including environmental microbiology, virology, neurobiology, molecular biology and immunology. ECL has improved the understanding and treatment of infectious diseases, cancer, neurodegenerative diseases and even sleep apnea disorders. Drug development has also benefited from ECL via improved assessment of pharmacodynamics, pharmacokinetics and determining immune responses against protein-based therapeutics. This review provides an overview of ECL chemistry and principles with a more detailed emphasis on the applications of ECL-based assays in different areas of science and medicine. The primary purpose of this review is to provide an in-depth discussion of the impact that ECL-based analysis has had on microbiology, immunology, virology, neurodegenerative diseases, molecular biology and drug development. Examples of ECL-based bioanalysis in each of these fields are discussed in conjunction with an overview of ECL principles and instrumentation.     

Genetics and genomics of infectious disease susceptibility

Human genetic variation is a major determinant of susceptibility to many common infectious diseases. Malaria was the first disease to be studied extensively and many susceptibility and resistance loci have been identified. However, genes for other diseases such as HIV/AIDS and mycobacterial infections are now being identified using a variety of approaches. A large number of genes appear to influence susceptibility to infectious pathogens and defining these can provide insights into pathogenic and protective mechanisms and identify new molecular targets for prophylactic and therapeutic interventions. Immunogenetic associations with infectious diseases have considerable potential to guide immunomodulatory interventions and vaccine design.     

Gut microbiota and the role of probiotics in therapy

Thanks to rapid progress in the development and application of molecular techniques to the assessment of the human gut microbiome, the true nature, diversity and metabolic potential of this 'hidden organ' are being revealed. Simultaneously, the complex physiological, immunological and metabolic interactions between host and microbiome are being untangled. By contrast, the probiotic concept has been with us for decades and, while supported more by fad and folklore in the past, is now gaining support, not only from experimental work in animal models, but also by well-designed studies in human diseases, most notably infectious diarrheas, inflammatory bowel disease and the irritable bowel syndrome.     

Approaches to minimize infection risk in blood banking and transfusion practice

The use of blood donor history and state-of-the-art FDA-licensed serological and nucleic acid testing (NAT) assays have greatly reduced the "infectious window" for several transfusion-transmitted pathogens. Currently transmission of human immunodeficiency virus (HIV), Human T-cell Lymphotropic Virus (HTLV), hepatitis viruses and West Nile Virus are rare events. The seroprevalence of cytomegalovirus in the donor population is high and cytomegalovirus infection can cause significant complications for immunocompromised recipients of blood transfusion. Careful use of CMV seronegative blood resources and leukoreduction of blood products are able to prevent most CMV infections in these patients. Currently, bacterial contamination of platelet concentrates is the greatest remaining infectious disease risk in blood transfusion. Specialized donor collection procedures reduce the risk of bacterial contamination of blood products; blood culture and surrogate testing procedures are used to detect potential bacterially contaminated platelet products prior to transfusion. A rapid quantitative immunoassay is now available to test for the presence of lipotechoic acid and lipopolysaccharide bacterial products prior to platelet transfusion. Attention has now turned to emerging infectious diseases including variant Creutzfeldt-Jakob disease, dengue, babesiosis, Chagas' disease and malaria. Challenges are presented to identify and prevent transmission of these agents. Several methods are being used or in development to reduce infectivity of blood products, including solvent-detergent processing of plasma and nucleic acid cross-linking via photochemical reactions with methylene blue, riboflavin, psoralen and alkylating agents. Several opportunities exist to further improve blood safety through advances in infectious disease screening and pathogen inactivation methods.     

Molecular epidemiology and evolution of emerging infectious diseases.

Molecular epidemiology is an emerging science. The development of new and rapid protocols to isolate and identify pathogens, coupled with the sophisticated phylogenetic analysis of their gene sequences, is providing a new and fascinating insight into the biology, origin and spread of infectious diseases. In this essay, I describe some of the ways in which the techniques of modern molecular biology and evolution have equipped us to face the challenge of these new infections.     

Nanotechnology and the diagnosis of dermatological infectious disease

Despite advances in diagnostics and therapeutics, infectious diseases continue to be a major cause of morbidity and mortality, surpassing cardiovascular diseases and cancer. Accurate identification of causative pathogens is critical to prevent the spread of infectious diseases and to deliver appropriate and timely therapy. Various limitations ranging from cost to lengthy yield times of current diagnostic modalities highlight the need for new approaches. Nanotechnology represents an innovative direction offering many advantages for pathogen detection and identification. Through surface modifications, nanoparticles can be tailored to bind microbial surface markers, nucleic acids, and toxins. Combining these nanoparticles with both standard and developing detection technologies has led to the development of faster, more sensitive, and more economical diagnostic assays. This review will focus on the diagnostic advances that utilize fluorescent, metallic, and magnetic nanomaterials, highlighting their potential applications in the diagnosis of infectious dermatological conditions.     

Molecular diagnosis of bloodstream infections caused by non-cultivable bacteria

Bloodstream infections are an important cause of morbidity and mortality in patients. Blood culture is clearly the most important diagnostic procedure for identifying micro-organisms involved in bloodstream infections except when the patient has previously received antibiotics or in the presence of slow-growing or intracellular micro-organisms. Detection of micro-organisms, mainly in blood, using pathogen-specific or broad-range PCR assays is promising. However, it is very important to emphasise that the interpretation of this molecular tool is critical because of the risk of interfering contamination, underlining the necessity to interpret the results obtained with caution. Presently, due to more widely available data and to rapid advances in biotechnology, two significant improvements allow new perspectives for molecular diagnosis. Indeed, the complete sequences of genomes have provided an important source of gene sequences for PCR-based assays. In addition, the development of real-time PCR offers several advantages in comparison to conventional PCR, including speed, simplicity, quantitative capability and low risk of contamination. Herein, we review the usefulness of molecular diagnosis of highly fastidious micro-organisms in the context of three different bloodstream infections: systemic diseases (rickettsiosis, Q fever, bartonellosis, Whipple's disease), blood-culture-negative endocarditis and bioterrorism attack.     

Rapid viral diagnosis by immunofluorescence reactions

Data are reviewed that illustrate the efficacy of the immunofluorescence (IF) technique as a method of rapid viral diagnosis. Its constant application in the "Stefan S. Nicolau" Institute of Virology has allowed the detection of viral and/or inframicrobial antigens in cells from patients with eye diseases (keratoconjunctivitis, keratitis, uveitis), stomatitis, skin diseases, benign and malignant gynecopathies, male urethritis, respiratory diseases, facial paralysis, etc. The IF technique has an incontestable advantage as regards the detection of the simultaneous presence of several infectious agents in the same patient. The prerequisites and prospects of a wider application of IF methods for rapid viral diagnosis are briefly discussed.     

Technology platforms for pharmacogenomic diagnostic assays

Rapid advances in the understanding of genomic variation affecting drug responses, and the development of multiplex assay technologies, are converging to form the basis for new in vitro diagnostic assays. These molecular diagnostic assays are expected to guide the therapeutic treatment of many diseases, by informing physicians about molecular subtypes of disease that require differential treatment, which drug has the greatest probability of effectively managing the disease, and which individual patients are at the highest risk of experiencing adverse reactions to a given drug therapy. This article reviews some of the relative strengths and limitations of the most widely used technologies and platforms for such assays.     

Point-of-care testing for infectious diseases: Opportunities,barriers, and considerations in community pharmacy

ObjectivesTo identify opportunities to perform point-of-care (POC) testing and/or screening for infectious diseases in community pharmacies, provide an overview of such tests and how they are used in current practice, discuss how the Clinical Laboratory Improvement Amendments of 1988 (CLIA) affect pharmacists performing POC testing, and identify and discuss barriers and provide recommendations for those wanting to establish POC testing for infectious diseases services in community pharmacies.Data sourcesPubMed and Google Scholar were searched from November 2012 through May 2013 and encompassed the years 2000 and beyond for the narrative review section of this article using the search terms rapid diagnostic tests, POC testing and infectious diseases, pharmacy services, CLIA waiver, and collaborative drug therapy management. All state boards of pharmacy in the United States were contacted and their regulatory and legislative websites accessed in 2012 and January 2013 to review relevant pharmacy practice laws.Data synthesisPOC testing for infectious diseases represents a significant opportunity to expand services in community pharmacies. Pharmacist education and training are addressing knowledge deficits in good laboratory practices and test performance and interpretation. Federal regulations do not define the qualifications for those who perform CLIA-waived tests, yet few pharmacists perform such services. Fewer than 20% of states address POC testing in their statutes and regulations governing pharmacy.ConclusionPOC testing for infectious diseases could benefit patients and society and represents an opportunity to expand pharmacy services in community pharmacies. Existing barriers to the implementation of such services in community pharmacies, including deficits in pharmacist training and education along with state regulatory and legislative variance and vagueness in statutes governing pharmacy, are not insurmountable.     

Hypertension 2020: confronting tomorrow's problem today

1. In developed countries, the major burden of disease is due to chronic diseases, such as heart disease and stroke. In contrast, the major burden of disease among people in developing countries has been due largely to diseases caused by malnutrition, poor sanitation and infection. In recent years, with increasing economic and demographic development, there has been a shift in developing countries from diseases caused by poverty towards chronic, non-communicable, lifestyle-related diseases. The rapid emergence of these chronic diseases has not occurred with a similarly rapid decline in infectious diseases. Therefore, these developing countries are experiencing high rates of both infectious and chronic diseases. 2. The increase in chronic diseases in developing countries has been brought about by the increasing prevalence of risk factors, such as increased alcohol consumption, smoking, obesity, physical inactivity and low fruit and vegetable intake. In parallel with this, there is also increased evidence of high blood pressure and high cholesterol levels. 3. The preventive strategies required to reverse this trend for these emerging diseases include the education of public health professionals, the introduction of surveillance activities to monitor changes in risk factors, the introduction of health promotion, the development of prevention research and improved advocacy for disease prevention programmes. Experience from other countries provides evidence that prevention programmes can work. The global challenge is to ensure that implementation of such programmes in the world's developing nations does not come too late.     

单克隆抗体在疾病治疗中的应用

单克隆抗体(单抗)技术与分子生物学技术的结合使重组嵌合抗体、人源化抗体和全人抗体的技术得到迅速发展.由于这些单抗带有人抗体的Fc片段,因此,这些抗体不仅能中和抗原,更重要的是能介导细胞毒作用和激活补体系统.这些抗体已被用于治疗某些疾病,如癌症、免疫性疾病、感染性疾病和器官移植排斥.随着大规模CHO细胞生产工艺的发展,越来越多的单抗将应用于临床.     

单克隆抗体在疾病治疗中的应用

单克隆抗体(单抗)技术与分子生物学技术的结合使重组嵌合抗体、人源化抗体和全人抗体的技术得到迅速发展.由于这些单抗带有人抗体的Fc片段,因此,这些抗体不仅能中和抗原,更重要的是能介导细胞毒作用和激活补体系统.这些抗体已被用于治疗某些疾病,如癌症、免疫性疾病、感染性疾病和器官移植排斥.随着大规模CHO细胞生产工艺的发展,越来越多的单抗将应用于临床.     

Technologies for individual genotyping: detection of genetic polymorphisms in drug targets and disease genes

Genetic variations have been associated with a predisposition to common diseases and individual variations in drug responses. Identification and genotyping a vast number of genetic polymorphisms in large populations are increasingly important for disease gene identification and pharmacogenetics. Commonly used gel electrophoresis-based genotyping methods for known polymorphisms include polymerase chain reaction (PCR) coupled with restriction fragment-length polymorphism analysis, allele-specific amplification, and oligonucleotide ligation assay. Fluorescent dye-based DNA fragmentation has been extensively used for high-throughput microsatellite or short tandem-repeat genotyping. TaqMan and molecular beacon genotyping are commonly used homogeneous solution hybridization technologies. Because of the ease of experimental assay design, single nucleotide polymorphism (SNP) genotyping methods based on single-base extension are in rapid development, such as fluorescence homogenous assays, pyrosequencing and mass spectrometry. Non-PCR based genotyping assays such as Invader trade mark assays are promised to genotype directly from genomic DNA without the requirement of PCR amplification. The DNA microarray is a solid phase genotyping format that is rapidly developing for parallel genotyping of a large number of SNPs simultaneously. Advanced technologies to identify genetic polymorphisms rapidly, accurately, and cost effectively will fundamentally change the practice of medicine by allowing physicians to prescribe medicine based on a patient's genetic make-up.