The development of a multiplex real-time PCR for the detection of herpes simplex virus 1 and 2, varizella zoster virus, adenovirus and Chlamydia trachomatis from eye swabs

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Abstract

Infectious conjunctivitis can be difficult to distinguish clinically due to the considerable overlap in clinical presentation so clinical diagnosis of conjunctivitis is often insufficient. It is therefore necessary to have a rapid diagnostic test that differentiates between the different causes of infectious conjunctivitis. Screening clinical samples by sample type/syndrome based multiplex real time PCR would allow for rapid detection of a variety of pathogens simultaneously, which will in turn aid in the treatment and clinical management of the patient.

A multiplex real-time PCR assay for rapid and simultaneous detection of HSV 1 and 2, VZV, adenovirus and Chlamydia trachomatis (C. trachomatis) from eye swabs was developed and evaluated. The multiplex assay was shown to be sensitive, specific and robust. Reductions in sample turn around times have been achieved by reducing the amount of separate tests needed to be carried out.

Highlights

► Infectious conjunctivitis can be difficult to distinguish clinically due to the considerable overlap in clinical presentation. ► A multiplex PCR assay for the screening of eye swabs was developed. ► This assay allows for the simultaneous detection of adenovirus, HSV 1 and 2, VZV and Chlamydia trachomatis in one reaction. ► Screening by multiplex real time PCR allows for rapid detection of pathogens simultaneously, which aids in treatment and clinical management. ► Since the introduction of the assay in 2007 the turn around time has reduced from 4.5 to 2 working days.

Abbreviations

PCR
polymerase chain reaction
Ct
cycle threshold
HSV
herpes simplex virus
VZV
varizella zoster virus

Keywords

Conjunctivitis
Viral conjunctivitis
Bacterial conjunctivitis
Multiplex real-time PCR
Eye swabs

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