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Elfath M. Elnifro, Robert J. Cooper, Paul E. Klapper, Adrian C. Yeo, Andrew B. Tullo; Multiplex Polymerase Chain Reaction for Diagnosis of Viral and Chlamydial Keratoconjunctivitis. Invest. Ophthalmol. Vis. Sci. 2000;41(7):1818-1822.
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purpose. To develop a multiplex polymerase chain reaction (PCR) for the
detection of adenovirus, herpes simplex virus, and Chlamydia
trachomatis in conjunctival swabs.
methods. Oligonucleotide primers for detection of the 3 agents were combined in
one reaction and evaluated for optimal performance using control DNAs
of adenovirus type 2, herpes simplex virus, and C.
trachomatis plasmid. The multiplex PCR was evaluated prospectively
against its corresponding uniplex PCRs, virus isolation, Chlamydia
Amplicor PCR, and an immunoassay technique (immune dot blot test) in a
total of 805 conjunctival swabs from patients with suspected viral and
results. The multiplex PCR was as sensitive as uniplex PCRs for the detection of
the agents in clinical specimens. In the prospective study, 48 of 49
(98%) clinical specimens were positive for adenovirus by the multiplex
PCR compared with 26 of 49 (53%) by adenovirus isolation. For herpes
simplex virus detection, the multiplex PCR had a sensitivity of 92%
(34/37) compared with 94.5% (35/37) by cell culture. The multiplex PCR
produced identical results to the Amplicor PCR (21/21; 100%) compared
with 71% (15/21) by the immune dot blot test.
conclusions. With clinical specimens the multiplex PCR was as sensitive as its
respective uniplex PCRs but more sensitive than adenovirus isolation
and as sensitive as herpes simplex virus isolation or C.
trachomatis Amplicor PCR. It has the potential to replace several
diagnostic tests with consequent savings in cost. The test also reduces
the risk of misdiagnosis by the clinicians.
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