Variplex™ test system fails to reliably detect SARS-CoV-2 directly from respiratory samples without RNA extraction

Eur J Clin Microbiol Infect Dis. 2020 Dec;39(12):2373-2377. doi: 10.1007/s10096-020-03983-9. Epub 2020 Jul 17.

Abstract

Diagnosis of COVID is performed by PCR methods, but their capacity is limited by the requirement of high-level facilities and instruments. The loop-mediated isothermal amplification (LAMP) method has been utilized for the detection of isolated virus-specific RNA. Preliminary data suggest the possibility of isothermal amplification directly from respiratory samples without RNA extraction. All patients admitted to our hospital were screened for SARS-CoV-2 by routine. Respiratory samples were tested by variplex system based on LAMP method directly without RNA extraction and by PCR. Primary endpoint was the false-negative rate of variplex test compared with PCR as gold standard. In 109 patients variplex test and PCR assay were performed simultaneously. Median age was 80 years and male/female ratio was 40/60%. The prevalence of PCR-confirmed COVID diagnosis was 43.1%. Variplex test was positive in 13.8%. False-negative rate of variplex test compared with PCR was 83.0%. The potential of LAMP technology using isolated RNA has been demonstrated impressively by others, and excellent sensitivity and specificity of detecting SARS-CoV-2 has been reported. However, without RNA extraction, the variplex test system failed to reliably detect SARS-CoV-2 directly in respiratory samples.

Keywords: COVID-19 diagnostic testing; False-negative reactions; Infectious disease outbreaks; Loop-mediated isothermal amplification; SARS-CoV-2; Specificity and sensitivity.

MeSH terms

  • Aged
  • Aged, 80 and over
  • Betacoronavirus / genetics*
  • Betacoronavirus / pathogenicity
  • COVID-19
  • COVID-19 Testing
  • Clinical Laboratory Techniques / methods*
  • Coronavirus Infections / blood
  • Coronavirus Infections / diagnosis*
  • Coronavirus Infections / epidemiology
  • Coronavirus Infections / mortality
  • False Negative Reactions
  • Female
  • Germany / epidemiology
  • Humans
  • Male
  • Molecular Diagnostic Techniques / methods*
  • Nasopharynx / virology
  • Nucleic Acid Amplification Techniques / methods*
  • Oropharynx / virology
  • Pandemics*
  • Pneumonia, Viral / blood
  • Pneumonia, Viral / diagnosis*
  • Pneumonia, Viral / epidemiology
  • Pneumonia, Viral / mortality
  • RNA, Viral / blood*
  • RNA, Viral / genetics
  • Reagent Kits, Diagnostic / standards
  • Retrospective Studies
  • Reverse Transcriptase Polymerase Chain Reaction / standards
  • SARS-CoV-2
  • Sensitivity and Specificity
  • Severity of Illness Index
  • Survival Analysis

Substances

  • RNA, Viral
  • Reagent Kits, Diagnostic

Supplementary concepts

  • LAMP assay