INFECTIOUS DISEASES
Strengthening Influenza Surveillance by Building Molecular Testing Capacity
By Alisa Bochnowski , senior specialist , Respiratory Diseases
APHL infecious disease team consultants and members of the Research Institute for Tropical Medicine , Philippines following Flu SC2 training
The Influenza Diagnostic Branch of the
US Centers for Disease Control and Prevention ( CDC ) developed a real-time multiplex PCR assay for the simultaneous detection of nucleic acid from influenza A , influenza B and SARS-CoV-2 viruses : the FluSC2 Multiplex . The assay was developed under Emergency Use Authorization ( EUA ) protocols to increase testing efficiency and to overcome laboratory constraints — such as molecular reagents and personnel shortages — which were globally experienced during initial
COVID-19 pandemic surge testing . By testing three major respiratory viruses that cause similar symptoms , the assay not only increases testing capacity for the laboratory but also contributes further to surveillance by understanding their trends and emergence . Because APHL was unable to host regional laboratory trainings due to the pandemic , and to support the implementation of the FluSC2 Multiplex Research Use Only ( RUO ) assay internationally , CDC led “ train-the-trainer ” sessions for APHL consultants on the FluSC2 Multiplex who subsequently deployed to different countries to provide support as the new assay was onboarded .
In January 2022 , CDC ’ s Influenza Genomics and Diagnostics Team , led by Dr . John Barnes , initiated training on FluSC2 Multiplex for a group of APHL influenza laboratory consultants .
The three-day hands-on training was conducted at CDC ’ s Atlanta campus and focused on the fundamentals of the assay and different troubleshooting techniques . Because not all CDC-validated devices are available internationally , the consultants also learned the RUO FluSC2 Multiplex verification process on other nucleic acid purification instruments and real-time PCR platforms including the Applied Biosystems QuantStudio™ 5 and the Bio-Rad CFX96 Real-Time PCR instruments .
To date , APHL has deployed consultants to Egypt , Jordan , Oman and the Philippines . Prior to deployment , APHL consultants worked with CDC and host laboratories virtually to ensure appropriate infrastructure and determine qualified personnel for the training . Upon arrival in country , the APHL consultants met with the laboratory director and personnel to provide an overview of the week . The consultants conducted a rigorous five-day training , including auditing lectures and assay setup and verification to ensure the assay can be performed optimally and reproduced in non-domestic settings . At the end of the training , the international laboratories were required to present their verification results ,
APHL infecious disease team consultants provided hands on technical assistance and training for the influenza and SARS-CoV-2 “ Flu SC2 ” multiplex PCR assay in Manila , Philippines
The EUA FluSC2 Multiplex assay is currently available to US state and local public health laboratories through CDC ’ s International Reagent Resource ( IRR ). The RUO FluSC2 Multiplex assay is available through IRR for international public health partners participating in the World Health Organization ( WHO ) Global Influenza Surveillance and Response Systems ( GISRS ) and other CDC-sponsored initiatives associated with pan-respiratory disease surveillance during the 2022 – 2023 influenza seasons .
provide an overall conclusion and receive additional feedback . Across the four in-country trainings , APHL consultants successfully trained more than 30 laboratorians from six different countries ( Egypt , Somalia , Iraq , Jordan , Oman and the Philippines ). These trainings allow participating countries to meet the demand of influenza and SARS-CoV-2 testing and will ultimately enhance global surveillance abilities . APHL and CDC hope to continue assisting countries through targeted technical assistance trips as well as regional trainings as global travel resumes . g
References :
1 . Shu , B , Kirby , MK , Davis , WG , Warnes , C , Liddell , J , Liu , J , et al . Multiplex Real-Time Reverse Transcription PCR for Influenza A Virus , Influenza B Virus , and Severe Acute Respiratory Syndrome Coronavirus 2 . Emerging Infectious Diseases , 2021 . 27 ( 7 ), 1821 – 1830 ( cited 2022 Sept 18 ). Available from
https :// doi . org / 10.3201 / eid2707.210462
22 LAB MATTERS Winter 2022