Clinical trial NCT03924284: Novel Assays for Detection of Influenza Virus
Official title: Evaluation of Novel Molecular Assays for the Detection of Influenza Virus
Study type: OBSERVATIONAL
Overall status: WITHDRAWN
Conditions: Influenza A Virus
Lead sponsor: The University of Hong Kong
Sponsor class: OTHER
First posted: 2019-04-23
Enrollment: 0
Sex: ALL
Standard ages: CHILD
ADULT
OLDER_ADULT
Accepts healthy volunteers: no
Observational model: OTHER
Time perspective: RETROSPECTIVE
Sampling method: NON_PROBABILITY_SAMPLE
Study population: These archived nasopharyngeal and saliva specimens are collected from patients in Queen Mary Hospital

Brief summary: Seasonal influenza virus causes an estimated 0.3-0.6 million deaths per year. Avian influenza virus H5N1, H7N9 and H5N6 has fatality rate of over 30%. Swine influenza viruses from pigs have also infected humans.

Molecular assays are now used routinely in the detection of influenza viruses. The M gene is often used as the target for all influenza A viruses because the nucleotide sequence of this gene is relatively conserved among all the influenza A viruses. The World Health Organization and the US Centers for Disease Control and Prevention (CDC) have published protocols for molecular detection of influenza A virus M gene.

However, recent studies have shown that mutations in the M gene have led to a reduced sensitivity of RT-PCR assay targeting this gene. Therefore, it is important to use alternative conserved genes as the target of RT-PCR. In this study, our aim is to evaluate two new RT-PCR assays that are based on PB2 and NS gene segment.

Detailed description: I. Background

* Seasonal influenza virus causes an estimated 0.3-0.6 million deaths per year. Avian influenza virus H5N1, H7N9 and H5N6 has fatality rate of over 30%. Swine influenza viruses from pigs have also infected humans.
* Molecular assays are now used routinely in the detection of influenza viruses. The M gene is often used as the target for all influenza A viruses because the nucleotide sequence of this gene is relatively conserved among all the influenza A viruses. The World Health Organization and the US Centers for Disease Control and Prevention (CDC) have published protocols for molecular detection of influenza A virus M gene.
* However, recent studies have shown that mutations in the M gene have led to a reduced sensitivity of RT-PCR assay targeting this gene. Therefore, it is important to use alternative conserved genes as the target of RT-PCR. In this study, our aim is to evaluate two new RT-PCR assays that are based on PB2 and NS gene segment

II. Study objective -To evaluate the sensitivity and specificity of 2 new RT-PCR assays

III. Overall study design

* The investigators will randomly retrieve archived nasopharyngeal and saliva specimens that were previously tested for influenza A virus using commercially available assays in our laboratory, tested for influenza A virus at the Public Health Laboratory Service Branch in Hong Kong. These specimens will be tested for influenza A virus by 4 different RT-PCR assays as listed below:

  1. Our new RT-PCR assay targeting PB2 gene
  2. Our new RT-PCR assay targeting NS gene
  3. M gene RT-PCR published by the World Health Organization
  4. M gene RT-PCR published by the US CDC

     Sensitivity, specificity, positive predictive value and negative predictive value will be determined.

     IV. Nucleic acid extraction and real-time reverse transcription-polymerase chain reaction (RT-PCR) for influenza A virus
* Saliva and nasopharyngeal specimens will be subjected to total nucleic acid (TNA) extraction by NucliSENS easyMAG (BioMerieux, Boxtel, Netherlands).
* Monoplex real-time RT-PCR assays for influenza A virus will be performed. The primers and probes for the M gene RT-PCR have been published by the WHO and the US CDC.

V. Sample size:

* The investigators will perform all 4 RT-PCR assays on a total of 320 specimens, including
* 80 nasopharyngeal specimens which tested positive for influenza A by commercially-available molecular assays or by testing performed at the Public Health Laboratory Services Branch in Hong Kong
* 80 nasopharyngeal specimens which tested negative for influenza A by commercially-available molecular assays or by testing performed at the Public Health Laboratory Services Branch in Hong Kong
* 80 saliva specimens which tested positive for influenza A by commercially-available molecular assays
* 80 saliva specimens which tested negative for influenza A by commercially-available molecular assays

Eligibility criteria: Inclusion Criteria:

* Nasopharyngeal or saliva specimens of patients in Queen Mary Hospital of Hong Kong
* Tested for influenza A virus using a commercially available assay or by the Public Health Laboratory Services Branch in Hong Kong

Exclusion Criteria:

* Insufficient specimen volume

Primary outcomes:
measure: RT-PCR result
timeFrame: Through study completion, an average of 2 months
Secondary outcomes:
measure: Cycle threshold value
timeFrame: Through study completion, an average of 2 months
Other outcomes:
Locations:
city: Hong Kong
country: Hong Kong
facility: Queen Mary Hospital
Study officials:
affiliation: The University of Hong Kong
name: Kelvin To, MD
role: PRINCIPAL_INVESTIGATOR
References: