Marietjie Venter1, Dewald Zaayman2, Stephanie van Niekerk2, Voula Stivaktas2, Shivani Goolab2, Jacqueline Weyer3, Janusz T Paweska3, Robert Swanepoel2. 1. Zoonoses Research Unit, University of Pretoria, Pretoria, South Africa; Centre for Respiratory Diseases and Meningitis, National Institute for Communicable Diseases of the National Health Laboratory Service, Sandringham, South Africa. Electronic address: marietjiev@nicd.ac.za. 2. Zoonoses Research Unit, University of Pretoria, Pretoria, South Africa. 3. Special Viral Pathogens Reference Laboratory, Centre for Emerging and Zoonotic Diseases, National Institute for Communicable Diseases of the National Health Laboratory Service, Sandringham, South Africa.
Abstract
BACKGROUND: Many cases of acute febrile illness with nervous manifestations go undiagnosed, partly because the potential pathogens are not investigated routinely. OBJECTIVE: To develop a multiplex PCR-based macroarray for detection of 29 pathogens associated with febrile disease, aseptic meningitis and meningoencephalitis in southern Africa, including common viruses, bacteria, parasites and selected arboviruses. STUDY DESIGN: Pathogens were identified by hybridization of PCR amplicons to probes on a macroarray chip, followed by colorimetric detection. RESULTS: Positive control specimens for all 29 targets were detected with high sensitivity. Twenty-seven clinical samples previously found positive for various etiologies of febrile disease and meningoencephalitis, including less common infections such as Crimean Congo haemorrhagic fever, Rift Valley fever, West Nile and rabies were all identified. Testing of a blinded panel of 16 specimens in triplicate demonstrated high repeatability. Screening of 138 specimens from patients with febrile and/or neurological signs that could not be solved in routine investigations yielded 5 additional diagnoses. CONCLUSION: A highly sensitive, specific and easily interpreted test was developed to detect 29 pathogens associated with febrile and nervous disease.
BACKGROUND: Many cases of acute febrile illness with nervous manifestations go undiagnosed, partly because the potential pathogens are not investigated routinely. OBJECTIVE: To develop a multiplex PCR-based macroarray for detection of 29 pathogens associated with febrile disease, aseptic meningitis and meningoencephalitis in southern Africa, including common viruses, bacteria, parasites and selected arboviruses. STUDY DESIGN: Pathogens were identified by hybridization of PCR amplicons to probes on a macroarray chip, followed by colorimetric detection. RESULTS: Positive control specimens for all 29 targets were detected with high sensitivity. Twenty-seven clinical samples previously found positive for various etiologies of febrile disease and meningoencephalitis, including less common infections such as Crimean Congo haemorrhagic fever, Rift Valley fever, West Nile and rabies were all identified. Testing of a blinded panel of 16 specimens in triplicate demonstrated high repeatability. Screening of 138 specimens from patients with febrile and/or neurological signs that could not be solved in routine investigations yielded 5 additional diagnoses. CONCLUSION: A highly sensitive, specific and easily interpreted test was developed to detect 29 pathogens associated with febrile and nervous disease.
Authors: Daisuke Mori; Wahida Khanam; Rahamot Ali Sheikh; S M Shahnawaz Bin Tabib; Emi Ikebe; Mohammad Moshaddeque Hossain; Hidekatsu Iha; Kamruddin Ahmed Journal: Sci Rep Date: 2017-11-23 Impact factor: 4.379