In studies of humoral immune responses to TB, the consensus is
that single antigen-based assays do not deliver satisfactory sero-
diagnostic performance. Combination tests that use several anti-
gens may be more useful for the development of diagnostic
antibody tests because of the large inter-individual variation in
serological responses. Many previous studies of new serodiagnostic
tests based on the well-known antigens have shown that combi-
nation of the speciﬁc antigen from M. tuberculosis gave higher
sensitivity than the individual antigen for assessing patients with
although not all the combinations always confer
This study showed that the sensitivity
was increased with the use of Rv3425, LAM plus the 38 kDa
antigen, and partly supported the above previous ﬁndings.
The present study has several limitations. First, serum samples
from healthy control subjects were all collected from Hebei Inter-
national Traveling Hygiene Health Center. Lack of the sera from
patients in whom pulmonary TB was initially suspected but who
were later found to have nontuberculous respiratory disease may
result in overestimate speciﬁcity of the multiple-antigen assay.
Second, clinical laboratory technicians performed TB-DOT and
TB-CHECK-1 tests not in a double blind manner, meaning they were
not blinded to patient diagnosis or results of the culture. However,
the multiple-antigen ELISA assays were conducted in a blinded way
by our investigators who were blinded to diagnosis and study
group. It would be better if all the tests were carried out in a blinded
In this study, our data suggest that the sensitivity and accuracy
of ELISA test based on the combination of Rv3425, 38 kDa and LAM
antigen are superior to those of single antigen-based assays and
those of two existing commercial kits. The higher sensitivity
and speciﬁcity, together with the simplicity and cost-effectiveness,
of the novel multiple-antigen ELISA test make it useful for the
diagnosis of smear-negative tuberculosis and of those subgroups of
patients with TB from whom specimens are difﬁcult to obtain i.e.
extra-pulmonary tuberculosis TB and childhood TB. Therefore, it
represents a promising tool for the serodiagnosis and screening of
active TB, particularly in developing countries and some areas with
high prevalence of TB infection and is worthy of further
This study was supported by China Mega-Projects of Science
Research for the 11th Five Year Plan (no. 2008ZX10003-003), China
Postdoctoral Science Foundation funded project (no.
20 080430624) and Shanghai Commission of Science and Tech-
nology, PR China (no. 08410703800).
Funding: Please see acknowledgements.
Competing interests: None declared.
Ethical approval: Not required.
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