Test system HSV 1 \x26amp; 2 IgG Plus – ZEUS Scientific

HSV Eraser Protocol
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Test system HSV 1 \x26amp; 2 IgG Plus – ZEUS Test system HSV 1 \x26amp; 2 IgG Plus An immunoassay based on microparticles and multiplexing for IgG antibodies to viral antigens Herpes Simplex 1 and 2 Product Number A97101G PROPER USE The AtheNA Multi-Lyte® HSV 1 \x26amp; 2 IgG Plus Zeus Scientific, Inc. aims qualitative detection of the presence or absence of IgG antibodies HSV-1 and HSV-2 in human serum. The test is indicated for sexually active adults and pregnant women, as an aid in the presumptive diagnosis of Herpes Simplex 1 and Herpes Simplex 2. The predictive value of positive or negative depends on the prevalence in the population and the likelihood of HSV 1 and HSV-2 before the test. The test is not intended to be used for the selection of donors or self-test. In performance of this assay has not been established for use in the pediatric population, immunocompromised patients, for use by ambulatory care facilities or for use with automated equipment. SIGNIFICANCE AND BACKGROUND Infections Herpes Simplex virus are caused by two distinct antigenic types, HSV-1 and HSV-2 (1). Both HSV types are common human pathogens. HSV-1 is usually associated with infections in the oropharyngeal area and eyes while HSV-2 causes most genital and neonatal infections (1,2). However, HSV-2 can occasionally isolated from the oropharyngeal area (3) and from 15% to 20% of primary genital infections may be caused by HSV-1 (1, 4). HSV infections are transmitted by virus containing secretions through close personal contact. HSV infections, both primary and recurrent are often subclinical and asymptomatic.

Excretion of the virus is the most important factor contributing to the spread of the virus (2). The primary HSV-1 infections of the oral mucosa usually occur in children under 5 years of age (2). Most infections are asymptomatic. Symptomatic infections are characterized by gingivostomatitis associated with fever, malaise and swollen cervical lymph nodes and soft (2). numerous small vesicles in the oral mucosa develop, ulcerate and heal in about two weeks. The most common form of recurrent HSV-1 is the herpes sores in which vesicles appear on the lips, nostrils or skin around the mouth (1, 2). Symptoms of genital HSV infections are accompanied by numerous ulcerative lesions pain, fever, and lymphadenopathy dysuria (6). The most severe complication of genital HSV infection is neonatal disease (2). In mothers with an active primary infection, the risk of transmission to infants can reach 40% (5). Approximately 69-80% of infants who develop neonatal herpes are born to women who are asymptomatic of genital HSV infection at the time of birth (5). Genital herpes is also problematic in sexually active adults and that often the disease is transmitted in the absence of symptoms (13). HSV antibody testing is indicated for sexually active adults to identify those at risk of acquiring or transmitting HSV HSV and other low risk of acquiring and transmitting infections HSV neonatal herpes (7,13) pregnant mothers. Although the combined culture with direct fluorescent antibody test (DFA) is definitive when making a diagnosis, timing and cultures should be obtained during periods of active disease to produce optimal recovery (8,9).

Serological procedures may be useful for diagnosis of primary HSV infections, and for determining evidence of past infection with HSV (10). However, many serological methods to determine HSV serostatus are unable to differentiate between HSV-1 and HSV-2 infections. Because the type of HSV implicated in disease has ramifications for prognosis (11. 12), it is important to specify the subtype. The specific serologic tests for HSV type were developed using the significant difference between the gG-1 protein gG HSV 1 and HSV-2 protein 2 (10). The first applications of specific serologic tests for HSV type HSV-1 and HSV-2 demonstrated that benefited tests gilts, recurrent and asymptomatic infections as a means of definitive diagnosis and counseling appropriate patient (13). The specific serological assays HSV type are useful in establishing or confirming the diagnosis of infections HSV-1 or 2 in asymptomatic persons, those with symptomatic lesions but negative culture, and those with atypical presentations (14). Specific tests the type of HSV are recommended for sexually active adults and pregnant women since the presence of antibodies to HSV is a reliable indicator that an individual can be infected with HSV and capable of transmitting the virus to others (14) . Assay principle AtheNA Multi-Lyte HSV Plus system AtheNA Multi-Lyte HSV 1 \x26amp; 2 IgG Plus Zeus Scientific, Inc. is a multiplexed assay designed to detect and differentiate IgG antibodies specific gG-1 and HSV gG-2 of HSV in human serum. The test procedure involves two incubation steps: 1. The patient sera were diluted and tested diluted sera are incubated in a vessel containing a multiplexed mixture of the microsphere suspension. The multiplexed bead suspension contains a mixture of distinguishable sets of polystyrene microspheres.

The antigen-specific gG-1 and HSV-2 gG protein are conjugated to the primary set of microspheres. The microsphere mixture also contains a bead set designed to detect non-specific antibodies in the patient sample (if present) and four separate bead sets used for assay calibration. If present in patient sera, HSV antibodies will bind to the corresponding set of microspheres immobilized antigens. The microspheres are rinsed to remove non-reactive serum proteins. 2. goat anti-human IgG conjugated with phycoerythrin (Fc chain specific) is added to the vessel and the plate is incubated. The conjugate will react with IgG antibody immobilized on the microspheres in step 1. Then mircroesferas suspension is analyzed by the AtheNA Multi- Lyte® instrument. The assembly or set of microspheres are sorted (identified) and the amount of reporter molecule (PE conjugate) is determined for each bead set. By using Intra-Well Calibration Technology® (intrapozuelo calibration technology), sets internal calibration bead is used to set the trial court. The raw fluorescence of each type of microsphere antigen gG-1 and HSV-2 gG is measured and compared relative to the calibrator in cutting. KIT COMPONENTS Reactive Reagents: (All reactive reagents contain sodium azide as a preservative at a concentration of 0. 1% w / v).

1. multiplexed bead suspension. Ready for use, 5. 5 mL bottle. The suspension contains polystyrene beads separate distinguishable 5. 6 micron conjugated with recombinant protein antigen gG-1 HSV-1, 55 kDa molecular weight and the recombinant protein antigen gG-2 HSV molecular weight 31 kDA. The microsphere mixture also contains a bead set designed to detect non-specific antibodies and / or IgM antibodies antireumatoide Factor (RF) in the patient sample (if present) in four separate bead sets used for calibration trial. 2. antihuman IgG conjugated with phycoerythrin goat (Fc specific chain). Ready to use, 15 mL amber bottle. Note: The bead suspension and conjugate are light sensitive reagents. Both have been packaged in containers protected from light. Normal exposures experienced during the course of performing the assay will not affect assay performance.

Do not expose these reagents to strong sources of visible light unnecessarily. 3. Control serum positive for HSV specific human HSV-1 and HSV-2. One, 0. 2 mL vial. 4. Control of human HSV negative serum. One, 0. 2 mL vial. 5. SAVe Diluent® (Diluent SAVe) A 50 mL bottle containing phosphate buffered saline. Ready to use. NOTE, the sample diluent will change color in the presence of serum.

6. Wash Buffer Concentrate: Dilute 1 part concentrate + 9 parts deionized water. A bottle containing 50mL mL 10X concentrated phosphate buffered saline. Nonreactive components: 1. A platen 96-well filtration for rinsing the microspheres. 2. Data Labels: One label is adhered to the inside lid of the kit box and a second label is inside the kit box. 3. Insert providing instruction packet for use. 4. CD calibration. A compact disc that includes the calibration values ​​specific kit batches required for specimen analysis and quality control testing. Materials required but not included: 1