Key result
The CkChpV-MIRA assay combined with a lateral flow dipstick detected CkChpV with a limit of 21.3 copies, demonstrating 100 times higher sensitivity than nested PCR without cross-reactivity.
Why the study?
Chicken chaphamaparvovirus is an emerging pathogen prevalent in chickens with diarrhea, prompting the need for more convenient and rapid diagnosis.
The CkChpV-MIRA assay provides a rapid, sensitive, and specific method for detecting chicken chaphamaparvovirus without requiring sophisticated equipment.
May enable rapid on-farm CkChpV detection; hypothesis-generating pending field validation in poultry.
Chicken chaphamaparvovirus (CkChpV) is a newly emerging pathogen that is currently prevalent in chickens with diarrhea symptoms. To diagnose CkChpV more conveniently and rapidly, this study established a multienzyme isothermal rapid amplification (MIRA) assay, with a reaction time of only 15 min and optimal reaction temperature of 38°C. In combination with the lateral flow dipstick assay, the CkChpV-MIRA assay can be completed within 20 min. We revealed that the detection limit of the MIRA assay using standard plasmids as templates was as low as 21.3 copies, and its sensitivity was 100 times higher than that of nested PCR. Moreover, the designed primer set and probe could only detect CkChpV specifically, and there was no cross reaction with avian nephritis virus, rotavirus, chicken parvovirus virus, Newcastle disease virus, and infectious bronchitis virus, which may cause diarrhea. These findings demonstrated that the CkChpV-MIRA assay established in this study is convenient, sensitive, and specific and does not require sophisticated equipment. It is more suitable for the detection of CkChpV in clinical samples.
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Cui et al. (2023) studied Chicken chaphamaparvovirus (CkChpV) infection. CkChpV-MIRA assay combined with lateral flow dipstick vs. Nested PCR was evaluated on Detection limit and sensitivity (100 times higher sensitivity). The CkChpV-MIRA assay combined with a lateral flow dipstick detected CkChpV with a limit of 21.3 copies, demonstrating 100 times higher sensitivity than nested PCR without cross-reactivity.
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