Researchers with Penn State and the USDA have developed a new test for early citrus greening detection.

Scientists with Penn State and the USDA have developed a test using CRISPR/Cas technology that could be used for citrus greening detection much earlier than anything else currently in use, according to a Citrus Industry article. Early detection of citrus greening is important to properly manage the disease. See the details below.

Technology for Early Citrus Greening Detection

Researchers have created a test using “the CRISPR/Cas system to develop an assay based on a platform known as DETECTR, which stands for DNA endonuclease-targeted CRISPR trans reporter. In this assay, a Cas variant called Cas12a is mixed with CRISPR RNA designed to seek out CLas DNA, and a synthetic molecule known as a fluorescent reporter oligonucleotide. If CLas DNA is present, the Cas12a cleaves both the pathogen’s DNA and the reporter oligonucleotide, enabling the generation of a fluorescent signal for detection,” according to the article.

They conducted a study with the test, recently published in the journal Phytopathology, and the test detected the citrus greening bacterium “at a sensitivity level 100 to 1,000 times greater than a commonly used diagnostic test, quantitative polymerase chain reaction (qPCR).”

Study co-author Yinong Yang said in the article “So far, various technologies, such as canine olfactory detection and qPCR, have been used to diagnose and confirm CLas infection. But these tools are often inadequate for early detection of CLas in asymptomatic tissue — where low amounts of bacteria are present — and are unsuitable for high-throughput diagnosis in the field.”

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