VDRL TEST: Venereal Disease Research Laboratory (VDRL) Test is a slide flocculation test employed in the diagnosis of syphilis.

Since the antigen used in this test is cardiolipin, which is a lipoidal e tracted from beef heart, it is not a specific test. This test is also classified as non!specific or non!treponemal or standard test. The antibodies reacting with cardiolipin antibodies ha"e been traditionally (but incorrectly) termed #regain$. Principle: %atients suffering from syphilis produce antibodies that react with cardiolipin antigen in a slide flocculation test, which are read using a microscope. &t is not 'nown if the antibodies that react with cardiolipin are produced against some lipid component of Treponema pallidum or as a result of tissue in(ury following infection. Requirements: %atient)s serum, water bath, freshly prepared cardiolipin antigen, VDRL slide, mechanical rotator, pipettes, and hypodermic syringe with unbe"eled needle and microscope. *nown reacti"e and non!reacti"e serum controls are also re+uired. VDRL antigen: The cardiolipin antigen is an alcoholic solution composed of ,.,-. cardiolipin, ,./0. lecithin and ,.1. cholesterol. The cardiolipin antigen must be freshly constituted each day of test. The wor'ing antigen is a buffered saline suspension of cardiolipin. VDRL slide: This is a glass slide measuring / 2 - inch with 0/ conca"e depressions, each measuring 03 mm in diameter and 0.45 mm deep. Procedure: %atients) serum is inacti"ated by heating at 53o6 for -, minutes in a water bath to remo"e non!specific inhibitors (such as complement). The test can be performed both +ualitati"ely and +uantitati"ely. Those tests that are reacti"e by +ualitati"e test are sub(ected to +uantitati"e test to determine the antibody titres. Qualitative test: ,.,5 ml of inacti"ated serum is ta'en into one well. 073,th ml (or 0 drop from 08 gauge needle) of the cardiolipin antigen is then added with the help of a syringe (unbe"eled) to the well and rotated at 08, rpm for 9 minutes. :"ery test must

be accompanied with 'nown reacti"e and non!reacti"e controls. The slide is then "iewed under low power ob(ecti"e of a microscope for flocculation. The reacti"e and non!reacti"e controls are loo'ed first to "erify the +uality of the antigen. Depending on the si;e the results are graded as wea'ly reacti"e (<) or reacti"e (R). Reacti"e samples are then sub(ected to +uantitati"e test. Qualitative test: This is performed to determine the antibody titres. The serum is doubly diluted in saline from 0in / to 0=/53 or more. ,.,5 ml of each dilution is ta'en in the well and 073, ml of antigen is added to each dilution and rotated in a rotator. The results are then chec'ed under the microscope. The highest dilution showing flocculation is considered as reacti"e titre. Sometimes, due to "ery high le"el of antibodies in the serum (pro;one phenomenon) the +ualitati"e test may be non! reacti"e. &f the clinical findings are strongly suggesti"e of syphilis, a +uantitati"e test may be directly performed on the serum specimen. CSF VDRL: VDRL test may also be performed on 6S> samples in the diagnosis of neurosyphilis. ?uantitati"e VDRL is the test of choice on 6S> specimens. @owe"er, there are some "ariations in this test. The antigen is diluted in e+ual "olumes with 0,. saline, 6S> must not be heated (or inacti"ated), the "olume of antigen solution ta'en is ,.,0 ml (or 0 drop from /0 gauge needle) and rotation time is 8 minutes. Rest of the procedure remains same. Significance of VDRL test: VDRL test becomes positi"e 0!/ wee's after appearance of (primary lesion) chancre. The test becomes reacti"e (5,!45.) in the late phase of primary syphilis, becomes highly reacti"e (0,,.) in the secondary syphilis and reacti"ity decreases (45.) thereafter. Treatment in the early stages of infection may completely suppress production of antibodies and result in non!reacti"e tests. :ffecti"e treatment in the primary or secondary stages results in rapid fall in titre and the test may turn non!reacti"e in few months. Treatment in latent or late syphilis has "ery little effect on the titre and the titres may persist at low le"els for long periods. Since the titre

falls with effecti"e treatment, it can be used for assessment of prognosis. VDRL test is more suitable as a screening agent than a diagnostic tool. VDRL test is also helpful in the diagnosis of congenital syphilis. Since passi"ely transferred antibodies through placenta may gi"e false reacti"e test in serum of the infant, a repeat test after a month showing no increase in titre may help rule out congenital syphilis. Since the test employs a non!treponemal antigen, there are many chances of false positi"e results. >alse positi"ity (Ather than technical) may be due to physiological of pathological conditions. These are called biological false positi"es (B>%). &f the remain positi"e for less than 3 months it is considered acute and they remain positi"e for longer than 3 months it is called chronic B>%. The physiological reasons for B>% include pregnancy, menstruation, repeated blood loss, "accination, se"ere trauma etc while the reasons for pathological B>% include malaria, infectious mononucleosis, hepatitis, relapsing fe"er, tropical eosinophilia, lepromatous leprosy, SL:, rheumatoid arthritis etc. C reacti"e VDRL test does not necessarily imply that the person is syphilitic. The diagnosis must be made in con(unction with clinical findings. Cny reacti"e VDRL test must be confirmed with a specific or treponemal test such as T%@C, >TC!CBS test.