The diagnostic method for Gastrodiscoides hominis relies on the microscopic examination of stool samples, a straightforward and commonly used method. No molecular (PCR or qPCR) diagnostic techniques are mentioned for Gastrodiscoides hominis; diagnosis is described only in terms of traditional parasitological stool microscopy.

"Gastrodiscoides hominis (Digenea: Paramphistomata: Gastrodiscidae) is an amphistomid intestinal fluke of pigs causing gastrodiscoidiosis. With the use of molecular tools assisting the conventional diagnostic procedures, we aimed at finding out molecular characterization of G. hominis using PCR amplifications of rDNA ITS (1, 2) sequences." The paper describes PCR amplification and sequencing of ITS1-2 rDNA regions for molecular characterization, but does not mention development of a quantitative (real‑time) PCR assay or its diagnostic validation in clinical or environmental samples.

In this paper, we report the design, development and validation of a novel *Gnomoniopsis castaneae* real‑time PCR detection assay, to our knowledge the first tool of this type available for this threatening pathogen. Thus, the aim of this work was to design and develop a highly sensitive and specific TaqMan assay for the detection of *G. castaneae* and submit it to a robust set of validation tests, including a comparison with parallel metagenomic data. The amplification efficiency of target DNA was 105.3% and the limit of detection of the assay was calculated at approximately 40 fg of pure fungal DNA, indicating that the assay was successfully developed and analytically validated, though directed at *G. castaneae* rather than *Gastrodiscoides hominis*.

A TaqMan quantitative PCR (qPCR) method assay targeting a novel genomic region of *A. cantonensis* was developed to detect infection in 150 snails collected from Hainan Province, China. The newly developed qPCR assay demonstrated superior sensitivity (reflected by lower Ct values) compared with previously published TaqMan qPCR methods. The established qPCR method provides a sensitive and non‑invasive tool for detecting *A. cantonensis* in snails, and can be applied for monitoring and early warning of parasite prevalence and transmission; however, this work does not involve *Gastrodiscoides hominis* and illustrates species‑specific TaqMan qPCR development for a different helminth parasite.

Here we report our development of a novel TaqMan qPCR assay based on the amplification of the conserved α‑tubulin gene fragment. The limits of detection, sensitivity and versatility of the α‑tubulin TaqMan assay were tested and validated using field samples from honeybee workers, wild bees, bumblebees and grasshoppers, as well as in the human infective trypanosomatid *Leishmania major*. These results show that the α‑tubulin TaqMan qPCR assay described here is a versatile diagnostic tool for the accurate detection and quantification of trypanosomatids in a wide range of environmental settings, demonstrating assay development and validation, but it is not specific to *Gastrodiscoides hominis*.

In this study, singleplex and duplex TaqMan qPCR were developed for the detection of the two viruses, based on two viral cloning vectors. A Cq value of ≤35 was employed as the Cq value determining the lower detectable limit of qPCR for the TuMV and BBWV2. The results demonstrate that, for both singleplex and duplex TaqMan qPCR assays, the lower detection limits are 2.38 × 10^1 copies/μL for BBWV2 and 1.12 × 10^3 copies/μL for TuMV, indicating consistent sensitivity across singleplex and duplex assays; this article illustrates assay development and validation methodology but concerns plant viruses rather than *Gastrodiscoides hominis*.

This study established a TaqMan probe‑based real‑time quantitative PCR assay targeting the small subunit rRNA (SSU) gene of *E. epinepheli*. The correlation coefficient (R^2) was 0.999, and the amplification efficiency was 106.4%. The detection limit of the TaqMan probe‑based qPCR assay was 1.0 × 10^1 copies/μL and that is 100 times more sensitive than the traditional PCR method. In addition, the test of clinical samples showed that this assay effectively detected *E. epinepheli* in the grouper's intestine tissue, demonstrating development and validation of a pathogen‑specific TaqMan qPCR assay, but the pathogen is a microsporidian, not *Gastrodiscoides hominis*.

Gastrodiscoides is a genus of zoonotic trematode under the class Trematoda. It has only one species, Gastrodiscoides hominis. Diagnosis is made by examination of the feces and the detection of eggs. Adult worms are easily identified from other helminths by their distinctive appearance. The eggs are readily distinguished from those of other trematodes by their rhomboid shape and distinct green colour. No reference is made to PCR or qPCR assays for the detection of Gastrodiscoides hominis DNA in clinical or environmental samples.

The eggs are very similar in size and appearance and difficult to distinguish morphologically from eggs of Fasciola hepatica and Gastrodiscoides hominis. The discussion in this CDC case study focuses on microscopic identification of trematode eggs and does not describe any quantitative PCR (qPCR) assays specifically targeting Gastrodiscoides hominis.

The China CDC Weekly article reports: "This study developed and validated an HT-qPCR assay capable of detecting 22 waterborne protozoa and helminths... Methods: We developed a high-throughput quantitative polymerase chain reaction (HT-qPCR) assay targeting 19 waterborne protozoa and 3 waterborne helminths and validated its sensitivity, specificity, and repeatability. The assay was then applied to test various environmental media samples." The list of targeted protozoa and helminths in tables and methods does not include Gastrodiscoides hominis, indicating that this particular environmental HT‑qPCR platform does not provide a specific assay for G. hominis DNA.

This study developed qPCR assay for Fasciola detection and validated a tremabiome deep amplicon sequencing method for fluke species differentiation. The work focuses on Fasciola and a panel of trematode species but does not list Gastrodiscoides hominis as a target, nor claim development of a qPCR assay specific for Gastrodiscoides hominis DNA in clinical or environmental samples.

Water tank experiments for in vitro validation of eDNA extraction and qPCR assay. Miracidia-containing water tank experiments and other tests were used to validate environmental DNA (eDNA) extraction and qPCR assay performance for trematode detection in water. The paper, however, does not describe assay development or validation specifically for Gastrodiscoides hominis; it focuses on other trematode species in environmental monitoring.

This study describes the development, optimisation and validation of a novel multiplex real-time PCR (qPCR) assay for the detection and differentiation of Cryptosporidium spp., Giardia duodenalis and Dientamoeba fragilis in clinical faecal samples. The assay achieved high diagnostic sensitivity and analytical sensitivity, including detection of the equivalent of a single Cryptosporidium oocyst. Gastrodiscoides hominis is not included among the targets of this multiplex qPCR assay.

"Here, we successfully developed a novel one-step multiplex qPCR assay that simultaneously identified and quantified L. martiniquensis and L. orientalis/L. chancei parasites and detected and quantified other trypanosomatids in clinical samples, using ITS1 as the molecular target and human RNase P as the internal control gene." The paper details assay design, analytical sensitivity/specificity and diagnostic performance in clinical DNA samples for several Leishmania and trypanosomatid species, but Gastrodiscoides hominis is not mentioned anywhere in the target list or discussion.

The article presents a "novel real‑time PCR assay based on the TaqMan probe" developed for the detection of *Gnomoniopsis castaneae*, the causal agent of brown rot of chestnut kernels. It states that the aim was "to design and develop a highly sensitive and specific TaqMan assay for the detection of *G. castaneae* and submit it to a robust set of validation tests, including a comparison with parallel metagenomic data". This is an example of TaqMan qPCR assay development and validation in plant pathology and does not concern *Gastrodiscoides hominis* specifically.

This report presents a highly sensitive real-time quantitative PCR (qPCR) assay developed to detect Blastocystis parasites in stool samples. A qPCR assay targeting a region of the Blastocystis small subunit rRNA gene was developed to detect and quantify subtypes ST1 to ST10 directly from human stool specimens, and the method was validated in epidemiological cohorts. The paper exemplifies qPCR assay development for intestinal parasites but does not mention Gastrodiscoides hominis or describe an assay for that species.

"This triplex qPCR method was shown to have good specificity, sensitivity, and reproducibility... In summary, triplex qPCR serves not only as a rapid and accurate detection and early prevention method for these pathogens but also constitutes a robust tool for microbial quality control in specific pathogen-free pigs." The assay targets Haemophilus parasuis, Streptococcus suis serotype 2, and Pasteurella multocida, with validation in clinical pig samples. No Gastrodiscoides hominis DNA detection or fluke-related assays are described, although pigs are the host species for G. hominis, suggesting that qPCR work in pigs to date has focused on bacterial pathogens rather than this trematode.

This article reports the validation of a quantitative PCR diagnostic method for detection of the microsporidian Ovipleistophora ovariae in cyprinid fish. The qPCR diagnostic method was validated using experimentally infected and naturally infected fish, demonstrating specificity and sensitivity for Ovipleistophora ovariae. Gastrodiscoides hominis is not discussed, indicating that validated qPCR assays exist for other parasites but not necessarily for Gastrodiscoides hominis.

This open-access review or case report on Gastrodiscoides hominis describes human gastrodiscoidiasis, clinical presentation, and treatment options. It emphasizes that diagnosis is typically made by detection of characteristic eggs in stool and does not mention the existence of a species-specific quantitative PCR (qPCR) assay validated for clinical or environmental detection of G. hominis DNA.

Gastrodiscoides hominis is a trematode fluke that primarily infects pigs and occasionally humans, with cases reported mainly from parts of Asia and Africa. Standard diagnostic approaches described in parasitology texts rely on microscopic identification of characteristic eggs in stool samples; there is no widely cited or standardised quantitative PCR (qPCR) assay in the clinical parasitology literature for Gastrodiscoides hominis as of 2024–2026.

This bioRxiv preprint describes development and validation of qPCR assays and deep amplicon sequencing for Fasciola species and broader trematode communities ("tremabiome"). The targets include Fasciola spp. and other trematodes relevant to livestock and humans, but Gastrodiscoides hominis is not listed as a species for which a specific qPCR assay was developed and validated.

This methods paper validates PCR-based protocols (including qPCR) for detection of Echinococcus multilocularis DNA in environmental samples such as fox feces and soil. It illustrates application of qPCR in parasitology and environmental detection of helminth DNA but is specific to Echinococcus and does not involve Gastrodiscoides hominis.

This paper validates an automated high-throughput multiplex real-time PCR panel for several intestinal parasites, including protozoa and helminths such as Giardia, Cryptosporidium, and some soil-transmitted helminths. Gastrodiscoides hominis is not included among the target pathogens, and the study does not present a qPCR assay specific to G. hominis.