Details

Autor(en) / Beteiligte

• Heiniger, Erin K.
• Buser, Joshua R.
• Mireles, Lillian
• Zhang, Xiaohong
• Ladd, Paula D.
• Lutz, Barry R.
• Yager, Paul

Titel

Comparison of point-of-care-compatible lysis methods for bacteria and viruses

Ist Teil von

• Journal of microbiological methods, 2016-09, Vol.128, p.80-87

Ort / Verlag

Netherlands: Elsevier B.V

Erscheinungsjahr

2016

Quelle

MEDLINE

Beschreibungen/Notizen

• Nucleic acid sample preparation has been an especially challenging barrier to point-of-care nucleic acid amplification tests in low-resource settings. Here we provide a head-to-head comparison of methods for lysis of, and nucleic acid release from, several pathogenic bacteria and viruses—methods that are adaptable to point-of-care usage in low-resource settings. Digestion with achromopeptidase, a mixture of proteases and peptidoglycan-specific hydrolases, followed by thermal deactivation in a boiling water bath, effectively released amplifiable nucleic acid from Staphylococcus aureus, Bordetella pertussis, respiratory syncytial virus, and influenza virus. Achromopeptidase was functional after dehydration and reconstitution, even after eleven months of dry storage without refrigeration. Mechanical lysis methods proved to be effective against a hard-to-lyse Mycobacterium species, and a miniature bead-mill, the AudioLyse, is shown to be capable of releasing amplifiable DNA and RNA from this species. We conclude that point-of-care-compatible sample preparation methods for nucleic acid tests need not introduce amplification inhibitors, and can provide amplification-ready lysates from a wide range of bacterial and viral pathogens. •Current methods of amplifiable nucleic acid recovery from pathogens remain too complex for simple point-of-care testing.•Lysis methods for bacteria and viruses need not introduce amplification inhibitors.•Different point-of-care-compatible lysis methods work best for different pathogen types.•The low-power AudioLyse device releases amplifiable DNA and RNA from Mycobacterium and Staphylococcus species.