The results were compared with those obtained from a commercial kit. Positive and negative sera for IBV were used as controls. To standardize the indirect ELISA development, serum dilutions (1:100, 1:200 and 1:400) and different concentrations of purified rN antigen (50, 100 and 200 ng/well) were tested. A total of 389 serum samples from chickens were used to develop and evaluate the ELISA-rN test. The rN of infectious bronchitis virus was purified by affinity chromatography using HisTrap HP 1 mL columns pre-packed with pre-charged Ni Sepharose in the ÄKTAprime Automated Liquid Chromatography system (GE Healthcare). coli BL21(DE3) Star competent cells (Invitrogen). MethodsĬonstructed recombinant pAE/ n expression vectors were used to transform E. The current study indicate that the combined use of rN in ELISA and WB is a powerful tool for the immunodiagnosis of avian infectious bronchitis. WB confirmed all false negative sera in ELISA-rN or IDEXX test as truly positive. ELISA-rN achieved a 90.34% sensitivity and 90.16% specificity. A total of 389 sera were tested against the rN in ELISA and the results were compared with those of the commercial IDEXX IBV Ab test. A soluble recombinant N protein (rN) of approximately 50 kDa was obtained. The coding sequence for N was amplified by RT-PCR and expressed in Escherichia coli. This study examined the combined use of an ELISA and Western blot (WB) to detect antibodies against the nucleocapsid protein (N) of IBV. Sometimes an antibody test for Lyme disease finds antibodies to other bacteria.The avian infectious bronchitis virus (IBV) remains a significant source of loss in the poultry industry and early diagnosis is required to prevent the disease from spreading. If Lyme disease bacteria DNA is found, you probably have an active Lyme disease infection.Once you have a Lyme disease infection, antibodies to the bacteria will usually stay in your body for the rest of your life. If antibodies are found, you may either have Lyme disease now or had the illness in the past.This is more likely with the antibody tests than with PCR.Īn abnormal, or positive, test for Lyme disease can mean one of the following: You have Lyme disease but it does not show up on the test ( false-negative).A negative PCR test usually means that you do not have a Lyme disease infection. A positive PCR test means the test detected Lyme disease bacteria DNA.Ī normal, or negative, test for Lyme disease can mean one of the following:.A negative PCR test means the test didn't detect Lyme disease bacteria DNA.The polymerase chain reaction (PCR) test may be done to confirm a Lyme infection if you have a positive antibody test result. An abnormal (positive) test means that Lyme disease antibodies are found.A normal (negative) test means that no Lyme disease antibodies are found.Test results are usually available in 1 to 2 weeks.Īn antibody test checks to see if you've had an infection from a bacteria that causes Lyme disease. The PCR test is a blood or urine test, but can also be done on fluid from the spine or from a joint. PCR testing may be used to identify a current (active) infection if you have symptoms of Lyme disease that have not gotten better with antibiotic treatment. Polymerase chain reaction (PCR) testing detects the genetic material ( DNA) of the Lyme disease bacteria. The Western blot test should be done for all people who've had a positive (or borderline positive) ELISA test. It is most often done to detect a chronic Lyme disease infection.Īntibody testing should be done in a two-step process, using the ELISA followed by the Western blot test. This test also identifies Lyme disease antibodies and can confirm the results of an ELISA test. This is a rapid test to identify Lyme disease antibodies. Enzyme-linked immunosorbent assay (ELISA).There are two types of antibody tests to detect Lyme disease. Finding antibodies to the Lyme disease bacteria does not tell whether you were infected recently or sometime in the past. Once formed, antibodies usually stay in your system for many years, even after successful treatment of the disease. It may take up to 2 months after becoming infected before antibodies can be detected in a blood test. Antibody testsĪntibody tests are the most commonly used tests to help identify Lyme disease. Other tests may be done in certain situations. Antibody tests can sometimes be used to help identify Lyme disease. If you and your doctor think you have Lyme disease, your doctor will do a careful medical history and physical exam. Lyme disease can be hard to diagnose because its symptoms are similar to those of many other illnesses. Lyme disease bacteria are spread by certain kinds of ticks. A Lyme disease test detects antibodies to the Lyme disease bacteria in the blood.
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