PCR: NAATs and RFLP

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The page below is a sample from the LabCE course Medically Important Aerobic Actinomycetes. Access the complete course and earn ASCLS P.A.C.E.-approved continuing education credits by subscribing online.

Learn more about Medically Important Aerobic Actinomycetes (online CE course)
PCR: NAATs and RFLP

Polymerase chain reaction (PCR) is a good method for identifying actinomycetes as it is based on the identification of specific genes. PCR tests use primers targeted at species-specific sequences.
  • So far, several PCR assays for Nocardia species, Streptomyces species, and Rhodococcus species have been developed for use in research.31 These types of nucleic acid amplification tests (NAATs) help to facilitate a rapid diagnosis.
    • Specific research targeted the rapid diagnosis of these three organisms directly from bronchoalveolar lavage (BAL) specimens of patients infected with HIV.32
    • PCR products were then sequenced to aid in obtaining full species identification.
PCR Restriction Fragment Length Polymorphism (PCR-RFLP molecular analysis [PRA]) involves the amplification of:
  • The hsp65 gene or the 16S rRNA gene, and
  • Subsequent digestions with specific restriction endonucleases for each gene31(p 20)
  • Useful for Nocardia isolates
31. National Health Service. Public Health England: UK standards for microbiology investigations - Identification of the aerobic actinomycetes. 2016. Accessed November 5, 2022. https://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/563971/ID_10i2.2.pdf
32. Rahdar HA, Salehi MR, Bahador A, et al. Detection of Nocardia, Streptomyces and Rhodococcus from bronchoalveolar lavage specimens of patients with HIV by multiplex PCR assay. Ethiop J Health Sci. 2019;29(6):737-744. doi:10.4314/ejhs.v29i6.10