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Loop-mediated isothermal amplification-based diagnostic assay for monkeypox virus infections.

Itoe Iizuka¹, Masayuki Saijo, Tomoyuki Shiota

¹Department of Virology 1, National Institute of Infectious Diseases, Musashimurayama, Tokyo, Japan.

Journal of Medical Virology

|April 22, 2009

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A new loop-mediated isothermal amplification (LAMP) assay rapidly and accurately diagnoses monkeypox virus (MPXV) infections. This sensitive diagnostic tool can differentiate between Congo Basin and West African MPXV strains.

Area of Science:

Virology
Molecular Biology
Infectious Diseases

Background:

Monkeypox virus (MPXV) causes a smallpox-like illness endemic to central and western Africa.
MPXV exists in two distinct genetic groups: the more virulent Congo Basin strain and the West African strain.
Accurate and rapid diagnostic methods are crucial for managing MPXV outbreaks.

Purpose of the Study:

To develop a real-time quantitative genome amplification system for MPXV diagnosis.
To utilize loop-mediated isothermal amplification (LAMP) technology for MPXV detection.
To create specific and multiplexed LAMP assays for differentiating MPXV strains.

Main Methods:

Designed primers targeting Congo Basin-specific (D14L gene), West African-specific (ATI gene), and shared (ATI gene) MPXV sequences.

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Developed Congo Basin (C-LAMP), West African (W-LAMP), and combined (COM-LAMP) real-time quantitative LAMP assays.

Evaluated LAMP assay sensitivity and specificity using nested PCR on primate blood and throat swab samples.

Main Results:

COM-LAMP demonstrated 80% sensitivity and 100% specificity; C-LAMP showed 79% sensitivity and 100% specificity; W-LAMP had 72% sensitivity and 100% specificity.
The LAMP assay accurately quantified MPXV viremia, correlating with disease severity.
The developed LAMP assays successfully discriminated between Congo Basin and West African MPXV strains.

Conclusions:

The novel LAMP assay is a rapid, quantifiable, highly sensitive, and specific system for MPXV diagnosis.
This LAMP assay enables differentiation between the two major MPXV genetic groups.
The developed LAMP technology is valuable for both diagnosing and assessing MPXV infections.