Development and validation of a multiplex PCR assay with melt curve analysis for detecting simian Plasmodium in wild Macaca fascicularis.

Thitiluck Swangsri, Rucksak Rucksaken, Wanat Sricharern, Wang Nguitragool, Naowarat Saralamba

Abstract readValidation Study

In one paragraph

Article in Scientific reports, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

0numbers the graph read from it

0cells of the map it votes in

0citing papers in PubMed

–field-weighted citation impact

1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

0 citing papers in PubMed.

No citing paper in PubMed yet.

4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

5 authors.

Thitiluck SwangsriDepartment of Molecular Tropical Medicine and Genetics, Faculty of Tropical Medicine, Mahidol University, Bangkok, 10400, Thailand.

Rucksak RucksakenDepartment of Veterinary Nursing, Faculty of Veterinary Technology, Kasetsart University, Bangkok, 10900, Thailand.

Wanat SricharernDepartment of Veterinary Nursing, Faculty of Veterinary Technology, Kasetsart University, Bangkok, 10900, Thailand.

Wang NguitragoolDepartment of Molecular Tropical Medicine and Genetics, Faculty of Tropical Medicine, Mahidol University, Bangkok, 10400, Thailand.

Naowarat SaralambaDepartment of Molecular Tropical Medicine and Genetics, Faculty of Tropical Medicine, Mahidol University, Bangkok, 10400, Thailand. naowarat.sar@mahidol.ac.th.

Funding

Mahidol University NSRF2024

6 · The paper itself

Abstract

Accurate and sensitive detection of simian malaria parasites is essential for surveillance and risk assessment of zoonotic malaria. We developed and validated a SYBR Green-based real-time PCR assay targeting the msp1 gene to detect and differentiate P. knowlesi, P. cynomolgi, and P. inui. Species-specific amplification was confirmed through distinct melting temperature (Tm) profiles. The assay demonstrated high analytical sensitivity, with a limit of detection of 10 copies/µL, excellent specificity with no cross-reactivity, and strong reproducibility, with low coefficients of variation for both cycle threshold (Ct) and Tm values. Amplification efficiency was within acceptable ranges, with R

Indexed as

Macaca fascicularisMalariaMultiplex Polymerase Chain ReactionPlasmodiumAnimalsMerozoite Surface Protein 1Real-Time Polymerase Chain ReactionReproducibility of ResultsSensitivity and SpecificityThailandTransition TemperatureMerozoite Surface Protein 1Macaca fascicularisMultiplex PCRPlasmodiumPlasmodium cynomolgiPlasmodium knowlesiReal-time PCR

Identifiers

PMID40595144

DOI10.1038/s41598-025-07337-3

PMCPMC12216036

What OpenQuestion holds

Textmetadata

LicenceCC BY

Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.