A Novel Nucleic Lateral Flow Assay for Screening phaR-Containing Bacillus spp

Area of Science:

• Microbiology
• Biotechnology
• Molecular Biology

Background:

• Polyhydroxyalkanoate (PHA) synthases are crucial for microbial PHA production.
• Class IV PHA synthases possess a unique PhaR subunit, encoded by the phaR gene.
• The phaR gene serves as a specific biomarker for identifying bacteria with class IV PHA synthases, including certain Bacillus species.

Purpose of the Study:

• To develop and validate a novel screening method for the phaR gene in Bacillus species.
• To enhance the specificity and sensitivity of phaR gene detection.
• To reduce the time and cost associated with screening for PHA-producing bacteria.

Main Methods:

• Development of a two-step screening method.
• Step 1: Polymerase Chain Reaction (PCR) for phaR gene amplification.
• Step 2: DNA lateral flow assay for phaR amplicon detection.

Main Results:

• The screening method demonstrated high specificity for phaR-containing Bacillus species.
• The method successfully detected as little as 10 pg of genomic DNA from B. thuringiensis ATCC 10792.
• The developed assay offers improved sensitivity and specificity compared to existing methods.

Conclusions:

• The novel screening method is a valuable tool for identifying phaR-containing Bacillus species.
• This method can be effectively applied to screen environmental samples for potential PHA producers.
• The improved screening process facilitates the discovery of superior PHA-producing microorganisms.