Key result
RT-LAMP matches PCR for HPeV3 detection but cuts diagnosis time to 2 hours.
Why the study?
Human parechovirus causes sepsis-like diseases and sudden infant death syndrome in neonates and young infants, creating a need for rapid and easier diagnostic tests.
Does the RT-LAMP technique allow for rapid and accurate detection of Human parechovirus in clinical samples?
Does the RT-LAMP technique allow for rapid and accurate detection of Human parechovirus in clinical samples?
Absolute Event Rate: 66.7% vs 66.7%
The newly developed RT-LAMP assay provides a rapid, easy, and specific method for diagnosing Human parechovirus infections in neonates and infants.
RT-LAMP matches PCR for rapid HPeV3 detection in small samples; hypothesis-generating and requires larger prospective validation before clinical adoption.
OBJECTIVES: Human parechovirus (HPeV), especially HPeV A3 (HPeV3), causes sepsis-like diseases and sudden infant death syndrome in neonates and young infants. Development of rapid and easier diagnostic laboratory tests for HPeVs is desired. METHODS: Original inner primers, outer primers, and loop-primers were designed on the 5' untranslated region of HPeV3. HPeV3 ribonucleic acids (RNAs), other viral RNAs, and clinical stool samples were used to confirm whether the designed primers would allow the detection of HPeV3 with the reverse transcription loop-mediated isothermal amplification (RT-LAMP) technique. RESULTS: Three combinations of primers were created and it was confirmed that all primer sets allowed the detection of HPeV3 RNAs. The primer sets had cross-reactivity with HPeV type 1 (HPeV1), but all sets showed negative results when applied to coxsackievirus, echovirus, enterovirus, norovirus, and adenovirus genomes. Four of six stool samples, obtained from newborn and infant patients with sepsis-like symptoms, showed positive results with our RT-LAMP technique. CONCLUSIONS: This manuscript is the first description of an RT-LAMP for the diagnosis of HPeVs, allowing a faster, easier, and cheaper diagnosis. This technique is clinically useful for newborns and infants who have sepsis-like symptoms.
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Yokoyama et al. (2021) studied Human parechovirus (HPeV) infection (n=6). Reverse transcription loop-mediated isothermal amplification (RT-LAMP) vs. Nested PCR was evaluated on Detection of HPeV3 in clinical stool samples. The newly developed RT-LAMP technique successfully detected HPeV3 in 4 out of 6 stool samples from infants with sepsis-like symptoms, matching nested PCR results but providing a diagnosis in 2 hours instead of 2 days.
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