Description

INTENDED USE
This EV-71 IgM ELISA is an enzyme-linked immunosorbent assay for qualitative determination of IgM-class antibodies to human enterovirus-71 (EV71)in serum or plasma samples. The assay is intended to be used in clinical laboratories for early diagnosis and management of patients related to infection with EV71.
SUMMARY
Human Enterovirus 71 (EV71), the newest member of Enterovirudae, is notable for its etiological role in epidemics of severe neurological diseases in children. It appears to be emerging as an important virulent neurotropic enterovirus in the upcoming era of poliomyelitis eradication. The illness usually peaks in June or July. EV71 infection may be asymptomatic or may cause diarrhea and rashes. EV71 one of the major causative agents for hand, foot and mouth disease (HFMD), is sometimes associated with severe central nervous system diseases.
Direct detection of virus is the mainstay of diagnosis. EV 71 can be isolated from throat and stool specimens, as well as from skin vesicle fuid. PCR testing provides generally greater sensitivity than culture for throat and stool specimens, and viral RNA has also been detected in vesicular fuid, blood and urine. EV 71-specifc serological assays, including tests specific for IgM antibody, have also been developed to assist for early and easier diagnosis of the disease.
PRINCIPLE OF THE TEST
This kit is a solid-phase antibody capture ELISA assay in which polystyrene microwell strips are pre-coated with antibodies directed to human immunoglobulin M proteins (anti-μ chain). The patient’s sample is added, and during the first incubation step, any IgM antibodies will be captured in the wells. After washing out all the other substances of the sample and in particular IgG-class antibodies, purified EV71 antigen, and monoclonal anti-EV71 antibodiesconjugated to the enzyme horseradish peroxidase (HRP-conjugate)are simultaneously added into the wells. Thise antigen and antibody will bind to any, previously captured onto the wells anti-EV71 antibodies. During the second incubation, the purified EV71 antigen and the HRP-conjugated antigens will specifically react only with the EV-71 IgM antibodies captured onto the wells during the first incubation. After washing to remove the unbound HRP-conjugate and excess EV71 antigen, chromogen solutions are added into the wells. In presence of EV-71 IgM antibody in the sample, the colorless chromogens are hydrolyzed by the bound HRP-conjugate to a blue-colored product. The blue color turns yellow after stopping the reaction with sulfuric acid. The amount of color intensity can be measured and is proportional to the amount of antibody captured in the wells, and to the amount of antibody in the sample respectively. Wells containing samples negative for EV71- IgM remain colorless.

In Vitro Diagnostic Use Only
UUUU | PLATE
Code5 (1x96wells)
8×12/12×8-well per plate MICROWELL PLATE: Blank microwell strips fixed on white strip holder. The plate is sealed in aluminium pouch with desiccant. Each well contains anti-IgM antibodies (anti-μ chain). The microwell strips can be broken to be used separately. Place unused wells or strips in the provided plastic sealable storage bag together with the desiccant and return to 2~8℃. Once open, stable for one month at 2-8℃
CONTROL | -
Code8 (1x0.5ml per vial)
preserv.0.1% ProClinTM 300 NEGATIVE CONTROL: Blue-colored liquid filled in a vial with green screw cap.
Protein-stabilized buffer tested non reactive for EV71- IgM.
Ready to use as supplied. Once open, stable for one month at 2-8℃
CONTROL | +
Code7 (1x0.5ml per vial)
preserv.0.1% ProClinTM 300 POSITIVE CONTROL: Red-colored liquid filled in a vial with red screw cap.
Purified EV71 IgM class antibodies diluted in protein-stabilized buffer.
Ready to use as supplied. Once open, stable for one month at 2-8℃.
bbDIL | SPE
Code9 (1x10ml per vial)
preserv.0.1% ProClinTM 300 SPECIMEN DILUENT: Blue liquid in a white vial with blue screw cap.
10ml per vial. Serum base, casein, and sucrose solution
Ready to use as supplied. Once open, stable for one month at 2-8℃.
bbAb | HRP
Code6 (1x6ml per vial)
preserv.0.1% ProClinTM 300
bbAg | REG
Code6A (1x6ml per vial)
preserv.0.1% ProClinTM 300 HRP-CONJUGATE: Red-colored liquid in a white vial with red screw cap.
Horseradish peroxidase-conjugated monoclonal anti-EV71 antibodies.
Ready to use as supplied. Once open, stable for one month at 2-8℃
ANTIGEN-REAGENT: Red-colored liquid in a white vial with blue screw cap.
Purified EV71 antigen.
Ready to use as supplied. Once open, stable for one month at 2-8℃
bWASH | BUF | 20X
Code1 (1x50ml per bottle)
DILUTE BEFORE USE! detergent Tween-20 WASH BUFFER: Colorless liquid filled in a clear bottle with white screw cap.
PH 7.4, 20 × PBS. The concentrate must be diluted 1 to 20 with distilled/ deionized water before use. Once diluted, stable for one week at room temperature, or for two weeks when stored at 2-8℃.
CHROM | SOL | A
Code 2 (1x6ml per vial) CHROMOGEN SOLUTION A: Colorless liquid filled in a white vial with green screw cap.Urea peroxide solution.
Ready to use as supplied. Once open, stable for one month at 2-8℃
CHROM | SOL | B
Code 3 (1x6ml per vial) CHROMOGEN SOLUTION B: Colorless liquid filled in a black vial with black screw cap.TMB solution (Tetramethyl benzidine dissolved in citric acid).
Ready to use as supplied. Once open, stable for one month at 2-8℃
,STOP | SOL
Code4 (1x6ml per vial) STOP SOLUTION: Colorless liquid in a white vial with white cap.
Diluted sulfuric acid solution (2.0M H2SO4).
Ready to use as supplied. Once open, stable for one month at 2-8℃
PLASTIC SEALABLE BAG, PACKAGE INSERT, CARDBOARD PLATE COVER