Chlamydia Trachomatis Antigen Detection by Chlamydiazyme Combined with Chlamydia Blocking Reagent Verification

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Abstract
Several options exist for the detection of chlamydial infection in a routine laboratory setting. Enzyme immuno assay (EIA) technology offers rapid turn around of results and is less technically demanding than chlamydial cell culture. In addition, recently introduced EIA confirmatory reagents have the potential to improve the accuracy of EIA detection. We have evaluated one such confirmatory reagent (Chlamydia Blocking Reagent®, Abbott Laboratories) to determine the accuracy of the Chlamydiazyme® EIA with special regard to interpretation of low absorbance values. An initial series of 192 male urethral specimens showed that use of a lowered cut off level (absorbance value 0.05) compared with that recommended by the manufacturer increased sensitivity of the EIA from 0.73 to 0.83, thus motivating studies on this interpretative modification. Of 1101 EIA reactive specimens, 65% were determined to be chlamydia positive by the Chlamydia Blocking Reagent. The proportion of female cervical specimens that did not confirm positive was elevated compared with male urethral specimens, 43% vs. 5.7% respectively. In samples yielding absorbance from the recommended cut off level to 0.05 (approximately 50% below), the corresponding figures were 78% and 14% respectively. In 85 selected EIA reactive samples, examination by a direct immunofluorescence staining assay (DFA) (MicroTrak®, Syva Inc.) revealed elementary bodies in 85% of 67 blocking test positive and in 24% of 18 blocking test negative samples. The possibility that Gram-negative bacteria were responsible for unconfirmed EIA reactive specimens was investigated using bacterial suspensions. While EIA reactivity was noted with several strains for Gram-negative bacteria, both the blocking reagent and DFA correctly verified the absence of chlamydial antigen. We conclude that confirming Chlamydiazyme initial reactive samples with the Chlamydia Blocking Reagent improved the accuracy of chlamydial detection by identifying false positive specimens. Furthermore, by including confirmatory testing, a lower cut off level than recommended by the manufacturer seems applicable, at least for male urethral samples, thus increasing sensitivity of the Chlamydiazyme EIA.

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