Immuno-assays - strategie used in diagnosis of human Infectious diseases

Abstract

Infectious diseases are a significant burden on public health and economic stability of societies all over the world. They have been among the leading causes of death and disability and presented growing challenges to health security and human progress for centuries. Infectious diseases are generally caused by microorganisms. The routes of them entry into host is mostly by the mouth, eyes, genital openings, nose, and the skin. Damage to tissues mainly results from the growth and metabolic processes of infectious agents intracellular or within body fluids, with the production and release of toxins or enzymes that interfere with the normal functions of organs and/or systems [1]. Advances in basic science research and development of molecular technology and diagnostics have enhanced understanding of disease etiology, pathogenesis, and molecular epidemiology, which provide basis for appropriate detection, prevention, and control measures as well as rational design of vaccine [2]. The diagnosis of infectious diseases is particularly critical for the prevention and control of the epidemic. Here we introduce the insights and detection methods of infectious disease, aiming to provide some helps for clinical diagnosis as well as epidemic prevention and control of infectious diseases.

Immuno-assays - strategie used in diagnosis of human Infectious diseases

The nucleic acid diagnostic tool currently employed is with good sensitivity and excellent specificity. However, due to its high false negative, time-consuming, high level equipment and technical personnel demand, the immunological antigen or antibody detection has been paid more and more attention because of its quick detection speed, low and simple technical requirements of detection. At present, the detection methods mainly include Enzyme-linked immunosorbent assays (ELISAs), colloidal gold immunochromatography (GICA) and magnetic particle chemiluminescence.

Enzyme-Linked Immunosorbent Assays (ELISA)

Enzyme-linked immunosorbent assays (ELISAs) incorporate the sensitivity of simple enzyme assays with the specificity of antibodies, by employing antigens or antibodies coupled to an easily-assayed enzyme. As such ELISA is much more rapid method than immunoblotting to detect specific viral protein from a cell, tissue, organ, or body fluid. There are two main variations of ELISAs: antigen-capture ELISA (detecting viral proteins), involve attachment of a capture antibody to a solid matrix for the viral protein of interest, while antibody-capture ELISA measures the specific antibody level in a sample, by coating viral antigen protein on a solid surface. There are two principles based on antigen-capture and antibody-capture ELISAs. In a general, ELISAs are considered a highly sensitive method that can detect a fairly low number of proteins at the range of picomolar to nanomolar range (10-12 to 10-9 moles per liter). ELISA has been one of the most widely used serologic tests for detecting antibody to HIV-1. ELISA method was found useful as a diagnostic tool to detect influenza viral antigen much quicker than other conventional virus detection methods [15]. In another previous study, comparison of ELISA, with conventional methods has demonstrated ELISA superiority for the rapid detection and identification of influenza A virus [16]. A simplified and standardized neutralization enzyme immunoassay (Nt-EIA) was developed to detect measles virus growth in Vero cells and to quantify measles neutralizing antibody [17]. Newer EIA formats for hepatitis C virus diagnostics have been constantly evaluated [18,19]. As such ELISAs are being used for plethora of application both in experimental and diagnostic virology including HIV-1, dengue, and influenza [20-22]. On the other hand, although rapid than traditional plaque assays or TCID50, ELISA assays sometimes could be quite expensive, due to the cost of reagents used. Unfortunately, sometimes required antibodies may not be commercially developed as well. In contrast, attempts to develop antibodies in-house may be quite expensive. Additional variability may also be introduced due to high background signals generated by non-specific binding, or cross-reactivity with non-viral protein targets.

Figure 2. A schematic representation of two principles based on antigen or antibody capture ELISA^[23] Colloidal gold immunochromatography (GICA)

Based on the specific immune response of antigen and antibody, colloidal gold particles were used as one of the tracer markers. Driven by solvent chromatography, the markers had an immune response on the C/T line, and the detection results could be obtained according to the color of the T line. GICA samples can be whole blood, serum or plasma, and studies have shown that the colloidal gold reagent has a high consistency in detecting whole blood, plasma or serum [24]. At present, there are seven kinds of colloidal gold kits approved by the State Food and Drug Administration, which are all detection antibodies, but there is no detection kit for antigens. RT-PCR was used as the control method, and the sensitivity and specificity of IgM/IgG antibody were different, and the highest detection rate of the two combined detection was 66.1% (125/189) [25]. This method can be used for enterprise resumption, students return to school, community crowd screening and other scenarios. Only a drop of fingertip blood is needed, and the detection results can be observed visually in 15 minutes, which is rapid and simple without special instruments. However, the detection has its disadvantages such as window period, without quantification, exposure risk, low sensitivity and vulnerability to environmental factors, and nucleic acid detection combination result is required for verification.

Figure 3. Scheme showing the general steps in the antibody-based diagnosis of viral infections from blood samples^[26] Magnetic particle chemiluminescence

Magnetic particle chemiluminescence is an emerging technique to capture specific IgM/IgG antibodies in samples using magnetic particles fixed on the surface of recombinant antigens. The antigen-antibody complex was precipitated by external magnetic field, and the captured antigen-antibody complex was identified by enzyme-labeled secondary antibody, and the luminescent intensity was determined by chemiluminescence instrument after adding the luminescent agent, and then quantitative analysis was carried out. It is characterized by high sensitivity, high specificity and wide detection range, etc. There are currently seven approved magnetic particle chemiluminescence detection kits, the first one developed by Bioscience, with an automatic chemiluminescence analyzer, capable of detecting at a speed of 240 T/H with an initial reporting time of 30 min [27].

Summary

Infectious diseases are a real public health threat, outbreaks can have serious social, political, and economic effects. A complex number of factors relating to human behavior and activities, pathogen evolution, poverty, and changes in the environment as well as dynamic human interactions with animals have been found to contribute to infectious disease emergence and transmission. Aggressive research is warranted to unravel important characteristics of pathogens necessary for diagnostics, therapeutics, and vaccine development. Here we describe some strategies for the diagnosis of human infectious diseases, hoping to be helpful for clinical diagnosis and epidemic prevention and control of infectious diseases. To date, multiple diagnostic techniques have been developed. Various diagnostic tools show both significances and limitations. Conventional approaches to quantify infective viral particles are labor-intensive, time-consuming, and often associated with poor reproducibility. Immunological tests generally provide quick results, however, is quite expensive due to the requirement of antigen-specific antibody. While RT-PCR may be able to provide results within a matter of hours, it is laborious, requires a skilled operator, and is sensitive to contamination. TEM-based quantification, although highly accurate in determining the shape and the total number of viral particles, often considered time-consuming, extremely expensive and impractical for high sample numbers. Moreover, TEM sample preparation is tedious, and the technique requires sophisticated instrument and a skilled operator. To alleviate these limitations, there is still a need to develop new cost-effective analytical methods that can allow users to quickly and easily determine virus concentrations and reduce constrictions coupled with current assays. Nevertheless, any such emerging methods must be carefully evaluated in terms of their efficiency, precision and linear range. The evaluation of each diagnostic technique and approval from the FDA are necessary before practical application.

Products list: paired diagnostics grade antibodies (monoclonal antibody, mab) and antigens for infectious disease rapid test kit

Cytokine release syndrome (CRS) is an acute systemic inflammatory syndrome characterized by fever and multiple organ dysfunction.

Genemedi produces core diagnostic ingredients for test of infectious diseases and related syndrome.

GeneMedi offers paired diagnostics grade antibodies (monoclonal antibody,mab)and antigens for infectious disease rapid test kit including infection of HIV/AIDS,Hepatitis C virus(HCV), Influenza A/B,Treponema Pallidum caused Syphilis,Helicobacter pylori (H. pylori) Bacteria, Herpes simplex virus 1/2(HSV-1/HSV-2),Cytomegalovirus (CMV),rubella virus,toxoplasma gondii and so on.

All the antibodies of inflammation and CRS test are suitable for in functional ELISA, and other immunoassays in diagnostics. The antibody can act as a capture antibody and detection antibody.

Classification: Astrovirus (Aastrovirus) human adenovirus anthrax bacillus acute respiratory infectious diseases Jejunum campylobacter candida Cytomegalovirus (CMV) Cryptosporidium (Crypto) parvum EB virus ebola virus (EV) Food source pathogenic bacteria resistant FluA FluB H. pylori Hepatitis b virus (HBV) HCV HIV1 HIV2 Human papilloma virus (HPV) herpes simplex virus HSV-1 HSV-2 Listeria monocytogenes Mycobacterium tuberculosis malaria Norovirus (NV, also called Winter Vomiting Bug) Prion Rift Valley Fever (RVF) rotavirus rubella virus salmonella Sexually transmitted diseases (STDs) TP(Treponema Pallidum) toxoplasma Vaccinia virus (cowpox virus) Vibrio cholerae West Nile virus (WNV)

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