Evaluation of BioPlex  2200 Syphilis IgG ®

for the diagnosis of syphilis

Marangoni A.1, Foschi C.1, Nardini P.1, Moroni A.2, Cevenini R.1,1Microbiology, DIMES, University of Bologna, 2U.O. Microbiology, St. Orsola Hospital, Bologna, Italy.

Introduction and Purpose

Syphilis is still a public health problem worldwide. The serological detection of specific antibodies to Treponema pallidum is
of particular importance in the diagnosis of the disease. Nowadays, most laboratories use reverse algorithm, especially when BioPlex® Syphilis IgG kit uses three different sets of beads coated with
huge numbers of sera need to be tested each day. The purpose of this study was to evaluate diagnostic performances of recombinant proteins from T. pallidum (15 KDa, 17 KDa, and 47 Kda). Data
BioPlex® 2200 Syphilis (BioRad), in comparison with ARCHITECT Syphilis TP (Abbott). As confirmatory tests, TPHA and Western are calculated as relative fluorescence intensity and converted to a
Blot (WB) were used. fluorescence ratio (FR) using an internal standard bead. The FR is
compared to an assay-specific calibration curve to determine the analyte
concentration in antibody index (AI) units and classified as negative
Methods
(<0.9), equivocal (0.9-1.1) or positive (>1.1).
In the Microbiology Laboratory of St. Orsola Hospital in Bologna a retrospective study was performed with the following five panels
of sera:

Group A: 100 samples from syphilis patients, with reactive RPR.

Group B: 100 samples from syphilis patients, with negative RPR. Specimens % reactivity
category (no. of samples reactive/total no. tested)
Group C: 53 samples reactive by Architect, but negative by TPHA, WB and RPR.
Group D: 100 samples from blood donors. BioPlex® ARCHITECT WB TPHA RPR
Syphilis IgG Syphilis TP
Group E: 200 samples from unselected patients submitted for serological syphilis screening.
100% 100% 100% 100% 100%
Group A (100/100) (100/100) (100/100) (100/100) (100/100)
All the 553 sera were analyzed by the following automated tests: BioPlex® 2200 Syphilis IgG (BioRad, Bio-Rad Laboratories,
Hercules, USA), an innovative test based on MFI technology (Multiplex Flow Immunoassay), and ARCHITECT Syphilis TP (Abbott,
Japan Co., Tokyo, Japan), a two-step immunoassay for the qualitative detection of IgG and/or IgM to T. pallidum using 100% 100% 100% 100% 0%
chemiluminescent microparticle immunoassay (CMIA) technology. Sera found reactive by at least one of the previous tests were Group B (100/100) (100/100) (100/100) (100/100) (0/100)
further analyzed by TPHA and RPR (Randox, Crumlin, UK), and “home made“ WB. A WB test was considered positive when at
least three bands out of TpN47, TmpA, TpN17 and TpN15 were clearly recognized. Titres ≥80 were considered as positive for 45,3% 100% 0% 0% 0%
TPHA testing. Group C (24/53) (53/53) (0/53) (0/53) (0/53)

Results 0% 0% 0% 0% 0%
Group D (0/100) (0/100) (0/100) (0/100) (0/100)
When used to define the immune response against T. pallidum in sera from groups A and B, both BioPlex 2200 Syphilis IgG and
®

ARCHITECT Syphilis TP showed excellent diagnostic performances compared to WB and TPHA findings (100% agreement). No
sample from group D was reactive when analyzed by all methods studied; moreover, all tests scored two samples among those 1% 1% 1% 1% 1%
from group E as reactive (100% agreement). Finally, BioPlex® 2200 Syphilis IgG scored as positive 23/53 sera of group C, whereas
Group E (2/200) (2/200) (2/200) (2/200) (2/200)
1/53 was Border line.

Conclusions
In the present study, specificity value of BioPlex® 2200 Syphilis IgG was higher than ARCHITECT Syphilis TP, when compared to WB and TPHA (93.2% and 84.9%, respectively), whereas sensitivity was 100%
for both tests. Because of its analytical performances, combined to BioPlex® system’s high throughput and walk-away capability, BioPlex® 2200 Syphilis IgG can be considered an excellent alternative to the
traditional treponemal tests.

Presented at the 23rd European Congress of Clinical Microbiology and Infectious Diseases, Berlin, 27-30 April 2013. (antonella.marangoni@unibo.it)