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Updated: Oct 11, 2025
Author Spotlight: Advancing Pathogen Diagnostics with Standardized LAMP
Published on: September 8, 2023
Rapid molecular diagnosis of Parechovirus infection using the reverse transcription loop-mediated isothermal
Tadafumi Yokoyama1, Yuko Tasaki1, Natsumi Inoue1
1Department of Pediatrics, Kanazawa University, Kanazawa, Ishikawa, Japan.
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.
Insights
A new reverse transcription loop-mediated isothermal amplification (RT-LAMP) technique rapidly detects Human parechovirus 3 (HPeV3). This faster, easier diagnostic method is crucial for identifying HPeV infections in infants with sepsis-like symptoms.
Area of Science:
- Virology
- Molecular Diagnostics
Background:
- Human parechovirus (HPeV), particularly HPeV A3 (HPeV3), is a significant cause of sepsis-like illnesses and sudden infant death syndrome in neonates and young infants.
- There is a critical need for rapid and accessible diagnostic tools for HPeV detection.
Purpose of the Study:
- To develop and validate a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for the rapid diagnosis of HPeV3.
- To assess the clinical utility of the developed RT-LAMP technique for HPeV detection in infant samples.
Main Methods:
- Novel inner, outer, and loop primers were designed targeting the 5' untranslated region of HPeV3.
- The RT-LAMP assay was optimized and tested using HPeV3 RNA, other viral RNAs, and clinical stool samples.
Main Results:
- All designed primer sets successfully detected HPeV3 RNA.
- The assay demonstrated specificity, with cross-reactivity only to HPeV1 and negative results for other enteroviruses and common respiratory/gastrointestinal viruses.
- Four out of six stool samples from infants with sepsis-like symptoms tested positive using the RT-LAMP technique.
Conclusions:
- This study presents the first RT-LAMP assay for HPeV diagnosis, offering a faster, simpler, and more cost-effective alternative to existing methods.
- The developed RT-LAMP technique is clinically relevant and valuable for the prompt diagnosis of HPeVs in newborns and infants presenting with sepsis-like symptoms.
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