CPHST: National Plant Pathogen Laboratory Accreditation Program Laboratory
CPHST Lab, Beltsville, Maryland
||Location: BARC-East, Bldg. 580, Powder Mill Rd., Beltsville, MD
Phone: (301) 313-9200
Fax: (301) 313-9232
Contact: Mark Nakhla
The Beltsville Laboratory mission is to develop, validate, and implement advanced biochemical and molecular methods for the detection of high consequence plant pathogens, including the APHIS Select Agents and plant pathogens in foreign germplasm. The CPHST laboratory now also conducts operational diagnostics of plant pathogen samples.
Laboratory programs utilize cutting-edge technologies from the fields of plant pathology, molecular biology, human and animal clinical diagnostics, and bio-detection to develop, adapt, and improve methods for accurate and rapid diagnosis of plant pathogens. The laboratory also diagnoses and differentiates high consequence and select agent plant pathogens that require federal confirmation. The Beltsville Lab strives to achieve timely transfer of diagnostic tools that are field deployable for PPQ emergency response and eradication programs. Tools are deployed to stakeholders through clearly written standard operating procedures and hands-on laboratory training for end users within and outside of PPQ.
The laboratory is a key component of the PPQ National Plant Pathogen Laboratory Accreditation Program (NPPLAP). Beltsville is responsible for proficiency test panel development, delivery, and first-level evaluation of proficiency tests conducted by scientists who perform diagnostics on behalf of PPQ using CPHST-validated methods. The laboratory is committed to quality in biochemical and molecular diagnostics, and is proficiency tested in the operation of diagnostic methods. Beltsville staff conduct outreach to the plant pathology diagnostic community by providing technical support to scientists within the National Plant Pathogen Diagnostic Network (NPDN), PPQ port and regional identifiers, and the state departments of agriculture in the detection of regulatory plant pathogens by providing protocols, hands-on laboratory training, and troubleshooting for PPQ validated diagnostics. Beltsville scientists also contribute their expertise by serving as members of scientific groups and committees.
- Developed a conventional PCR assay for the for the detection of the Nepovirus Subgroup B for germplasm screening.
- Characterized a novel virus causing symptoms similar to citrus leprosis and proposed the virus be called Citrus leprosis virus cytoplasmic type 2.
- Evaluated an ELISA test for detection of Citrus leprosis virus-C.
- Developed assays to differentiate the sweet orange fungal pathogen, Elsinoe australis subgroups and species.
- Developed conventional and quantitative PCR molecular diagnostics for the late wilt of corn fungal pathogen, Harpophora maydis.
- Developed molecular markers for the differentiation of isolates of Phytophthora kernoviae from Great Britain and New Zealand.
- Developed molecular assays for the detection and differentiation of pathogenic nematodes Anguina funesta, A. agrostis, A. tritici, and A. pacificae. Morphological identification is difficult and labor intensive, and these assays provide a simpler method to differentiate these species.
- Validated real-time PCR methods for detection and identification of Rathayibacter toxicus, a toxigenic bacterium vectored by the nematode Anguina funesta, that can cause disease in livestock that eat infected ryegrass.
- Developed assays to detect two additional genes of Candidatus Liberibacter asiaticus associated with citrus Huanglongbing (HLB).
- Evaluated and adapted CANARY technology for rapid detection of Phytophthora. The optimized Phytophthora CANARY system was used to successfully detect field collected and laboratory inoculated samples.
- Evaluated and adapted the Lincoln Nucleic acid Kit (LiNK) technology for rapid extraction of plant pathogen DNA. The LiNK sampling methods were approximately 100 times more sensitive than the Qiagen DNeasy Plant Mini Kit with a processing time of 6 minutes vs. 1 hour for the Qiagen Kit. This system will be further optimized for use in plant pathogen diagnostics.
- Prepared proficiency test panels for HLB/citrus greening, Plum pox virus, and Phytophthora ramorum or the PPQ National Plant Protection Laboratory Accreditation Program.
- Completed and released 9 new or revised work instructions documents. These covered a diverse range of diagnostic methods, training instructions, and DNA/RNA isolation protocols.
- Provided three different hands-on training sessions that met on five total occasions for 42 scientists and diagnosticians from PPQ, NPDN, and universities.
- In fiscal year 2012, completed regulatory diagnoses for 13 different plant pathogens. A total of 1606 individual samples were tested.