Multiplex PCR For Detection Research Articles

Multiplex PCR detection of enteric pathogens in a community-based birth cohort in Ecuador: comparison of xTAG-GPP and TaqMan array card assays.

We compared the performance of two multiplex platforms, Luminex xTAG Gastrointestinal Pathogen PanelⓇ and TaqMan Array Card, against a panel of 14 enteric pathogen targets in a community-based birth cohort in Ecuador. We found high levels of agreement and similar prevalence estimates across most pathogens.

A rapid, sensitive, and high-throughput pathogen detection method and application of identifying pathogens based on multiplex PCR technology combined with capillary electrophoresis

Respiratory infections are one of the leading causes of death worldwide. Rapid and accurate detection of pathogens is crucial for timely treatment. This study established a detection method based on multiplex PCR combined with capillary electrophoresis, capable of simultaneously detecting 28 common respiratory pathogens. We used specially designed homo-tag primers, significantly reducing the formation of primer dimers and improving the specificity and sensitivity of amplification. After two rounds of PCR amplification, the products were subjected to fragment analysis using the ABI 3500XL Genetic Analyzer, enabling automated result interpretation. The detection limit of this method reaches 2.77 × 102 copies/ml, with some pathogens reaching as low as 2.77 × 10 copies/ml. The detection of 147 clinical specimens showed that the overall consistency of this method with culture, colloidal gold, fluorescent quantitative PCR, and NGS was 75.5%. The multiplex PCR detection method established in this study is rapid, sensitive, and high-throughput, can be used for routine screening of common pathogens in respiratory infections, providing an important basis for clinical diagnosis and treatment.

Multiplex-PCR Detection of Clostridium tyrobutyricum, Clostridium butyricum, and Clostridium sporogenes in Raw Milk for Cheesemaking.

Late blowing defects in semi-hard and hard cheeses caused by spore-forming clostridia (e.g., Clostridium tyrobutyricum, Clostridium butyricum, Clostridium sporogenes) pose a major issue for the dairy industry. With this study, we applied a multiplex PCR for the rapid and simultaneous detection of clostridia in raw milk for cheese production. Spore detection in milk usually relies on culture-dependent methods, among which the most probable number (MPN) technique is sensitive but time-consuming and nonspecific. We tested two PCR-based protocols: the one entailed direct milk analysis with results obtained within 24 h; the other included an enrichment step and gave results within 72 h. The second protocol was found to be more sensitive; it detected concentrations as low as 100 cells/L for C. sporogenes and C. butyricum and 800 cells/L for C. tyrobutyricum. Both protocols were applied to field samples (211 samples underwent protocol no. 1; 117 samples underwent protocol no. 2) collected from four dairy processing plants in Piedmont. The prevalence of C. butyricum (protocol no. 1: 9.5%; protocol no. 2: 23%) was higher than either C. sporogenes (0%; 9.4%) or C. tyrobutyricum (0%; 6.8%). Protocol no. 2 detected multiple targets in eight samples, indicating that more than one microorganism was present. Our findings underscore the importance of implementing preventive measures and early detection strategies to mitigate the risk of cheese spoilage due to clostridial contamination.

Multiplex PCR assay for the accurate and rapid detection and differentiation of Lactococcus garvieae and L. petauri.

Lactococcosis is a common bacterial fish disease caused by Lactococcus garvieae, L. petauri and L. formosensis. Although there are different PCR-based techniques to identify the etiological agent, none of these can differentiate these two bacteria without sequencing PCR-amplified fragments. In the present study, we developed a multiplex PCR assay for simultaneous detection and differentiation of L. garvieae and L. petauri. The specificity of the primers was validated against the bacterial DNA of the targeted and non-targeted bacteria. The sizes of the PCR amplicons were obtained as 204 bp for the DUF1430 domain-containing protein gene of L. garvieae, 465 bp for the Lichenan permease IIC component gene of L. petauri, and 302 bp for the teichoic acid biosynthesis protein F gene of both L. garvieae and L. petauri. The PCR amplicons were clearly separated by agarose gel electrophoresis. The multiplex PCR assay did not produce any amplification products with the DNA of the non-targeted bacteria. The multiplex PCR detection limits for L. garvieae and L. petauri were 5 and 4 CFU in pure culture and 50 and 40 CFU/g in spiked tissue samples, respectively. It takes less than 2 h from plate-cultured bacteria and 3 h from tissue samples to get results. In conclusion, the developed multiplex PCR assay is a rapid, specific, accurate, and cost-effective method for the detection and differentiation of L. garvieae and L. petauri and is suitable to be used for routine laboratory diagnosis of L. garvieae and L. petauri.

Multiplex-PCR Detection of an Atypical Leuconostoc mesenteroides subsp. jonggajibkimchii Phenotype Dominating the Terminal Spoilage Microbial Association of a Fresh Greek Whey Cheese Stored at 4 °C in Vacuum

Multiplex-PCR Detection of an Atypical Leuconostoc mesenteroides subsp. jonggajibkimchii Phenotype Dominating the Terminal Spoilage Microbial Association of a Fresh Greek Whey Cheese Stored at 4 °C in Vacuum

Establishment and Validation of a Multiplex PCR Detection System for the Identification of Six Common Edible Meat Components.

To establish a rapid, accurate, and sensitive multiplex PCR detection method for the simultaneous identification of the six common edible meats (beef, lamp, chicken, pork, goose, duck), and to evaluate its application value in meat adulteration identification. Based on complete mitochondrial genomic sequences of six species in the GenBank database, DNA sequences (cattle：16S rRNA; sheep：COX-1; chickens：Cytb; pig：COX-1; goose：NADH2; duck：16S rRNA) with intra-species conservation and inter-species specificity were screened, and species-specific primers were designed to construct a multiplex PCR detection system that can simultaneously detect the meat of six common species. The species specificity, sensitivity and reproducibility of the system were studied, and the simulated mixture sample detection was performed. This study successfully constructed a multiplex PCR detection system that can detect the meats of six common species simultaneously. The system was not effective in DNA amplification of non-target species. When the DNA template sizes were 0.062 5-2 ng/μL, the amplified products of all six species could be detected. The duck component was still detected when the mixing ratio of duck and beef was as low as 0.5%. This study constructs and establishes a multiplex PCR detection system with strong specificity, high sensitivity, and good reproducibility. It can accurately identify the components of animal origin in common edible meats and provide a simple and practical method for identifying adulteration of common edible meats and meat products in China.

The development of novel genome-SSRs, multiplex PCR panels, and allelic ladders for parentage identification in Tachypleus tridentatus

Tachypleus tridentatus, one of the oldest animal species on Earth, with significant biological and ecological value, its population sizes have declined due to overfishing. Stock enhancement technology has been used to protect T. tridentatus, however, its efficiency needs to be addressed. In the present study, a parentage identification system was developed. We identified 235,336 SSRs and 182,394 primer pairs were designed. Out of 321 synthesized primers, 14 highly polymorphic primers were selected to reveal the genetic diversity of 110 T. tridentatus. SSRs were amplified with the 14 developed primers, which were fluorescently labeled, and the resulting PCR products were analyzed using the ABI DNA Analyzer 3730xl. The number of alleles per locus, the polymorphism information content, the observed heterozygosity, and the expected heterozygosity ranged from 7 to 48, 0.581 to 0.963, 0.636 to 0.909, and 0.645 to 0.969, respectively. The cumulative exclusion probability with both unknown parents, with only unknown parents, or with both known parents was >99.9999% for the 14 loci. Furthermore, to facilitate data exchange across laboratories and platforms, multiplex PCR detection, SSR fragment size ladders, and parentage identification were employed. Out of the 14 primers, 6 optimal primers were selected for multiplex PCR validation, and two sets of triplex systems were established. Three thousand clones for 160 alleles of the six primers were selected for plasmid extraction and sequencing, and 600 correct plasmids were obtained for new T. tridentatus SSR fragment size ladder construction. Two pairs of parents with 12 offspring each were used for parentage identification by the multiplex PCR systems and ladders, the cumulative average exclusion probability of these 6 primers reached 99.9999% among parents and their offsprings, and was 0% in nonparent-offspring group. The parentage identification system constructed in this study can serve as a tool for monitoring the survival rates of T. tridentatus in stock enhancement programs.

Development of a multiplex real-time PCR assay for the simultaneous detection of mpox virus and orthopoxvirus infections

Since May 2022, the multi-country outbreak of monkeypox (mpox) has raised a great concern worldwide. Early detection of mpox virus infection is recognized as an efficient way to prevent mpox transmission. Mpox specific detection methods reported up to now are based on the SNPs among mpox virus and other orthopoxviruses. We have therefore developed a real-time PCR based mpox detection method targeting mpox virus specific sequences (N3R and B18Rplus). We have also optimized an orthopoxvirus detection system which targets the highly conserved E9L and D6R genes. The mpox and orthopoxvirus real-time PCR assays have a high sensitivity (1 copy/reaction) and specificity. Mpox viral DNA and clinical samples from mpox patients are detected with the mpox detection system. Furthermore, we have established a multiplex real-time PCR detection system allowing simultaneous and efficient detection of mpox and orthopoxvirus infections.

Development and preliminary application of a quadruplex real-time PCR assay for differential detection of porcine circovirus 1-4 in Chengdu, China.

Porcine circovirus (PCV) typically causes severe immune suppression in pigs, leading to mixed clinical infections with various pathogens that can cause significant harm to the pig industry. PCV has four subgenotypes, with PCV4 being an emerging virus that requires investigation due to its potential for epidemic outbreaks. Therefore, there is a need to develop a method that can detect all four PCV strains simultaneously. In this study, four pairs of specific primers and TaqMan probes were designed based on the conserved sequence of the PCV1-4 ORF2 gene to establish a PCV1-4 TaqMan multiplex real-time quantitative PCR method. The novel method was compared to six commercial testing kits for its efficacy. Then, a total of 595 mixed samples of spleen and lymph node collected from 12 districts in Chengdu from July to December 2021 were tested using the novel method. The results showed that the novel PCV1-4 TaqMan multiplex real-time quantitative PCR detection method has satisfied specificity, sensitivity, and repeatability. The positive rates of PCV1, PCV2, and PCV3 in Chengdu were 2.18%, 31.60%, and 15.29%, respectively, while no positive PCV4 was detected. The mixed infection rate of PCV2 and PCV3 was 5.21%. Our novel method may be as a potential method for PCV1-4 detection. Currently, PCV2 is the main epidemic PCV subtype in Chengdu, while the potential threat of PCV4 should also be considered.

Occurrence and Multiplex PCR Detection of Citrus Yellow Vein Clearing Virus in Korea.

Citrus yellow vein clearing virus (CYVCV) is a member of the Alphaflexiviridae family that causes yellow vein clearing symptoms on citrus leaves. A total of 118 leaf samples from nine regions of six provinces in Korea were collected from various citrus species in 2020 and 2021. Viral diagnosis using next-generation sequencing and reverse transcription polymerase chain reaction (RT-PCR) identified four viruses: citrus tristeza virus, citrus leaf blotch virus, citrus vein enation virus, and CYVCV. A CYVCV incidence of 9.3% was observed in six host plants, including calamansi, kumquat, Persian lime, and Eureka lemon. Among the citrus infected by CYVCV, only three samples showed a single infection; the other showed a mixed infection with other viruses. Eureka lemon and Persian lime exhibited yellow vein clearing, leaf distortion, and water-soak symptom underside of the leaves, while the other hosts showed only yellowing symptoms on the leaves. The complete genome sequences were obtained from five CYVCV isolates. Comparison of the isolates reported from the different geographical regions and hosts revealed the high sequence identity (95.2% to 98.8%). Phylogenetic analysis indicated that all the five isolates from Korea were clustered into same clade but were not distinctly apart from isolates from China, Pakistan, India, and Türkiye. To develop an efficient diagnosis system for the four viruses, a simultaneous detection method was constructed using multiplex RT-PCR. Sensitivity evaluation, simplex RT-PCR, and stability testing were conducted to verify the multiplex RT-PCR system developed in this study. This information will be useful for developing effective disease management strategies for citrus growers in Korea.

Virus removal by high-efficiency air (HEPA) filters and filtration capacity enhancement by nanotextiles: a pilot study.

Portable household air purifiers are widely used devices designed to maintain a high-quality level of indoor air. Portable air purifiers equipped with the high-efficiency air (HEPA) filter served 100h in a household space occupied by two adults without any symptoms of respiratory tract infection. The main objective of the study was to determine microbial contamination on the HEPA filter and to investigate if the selected nanotextile monolayer made of polyamide 6 (PA6) nanofibers can capture potential microorganisms when installed downstream of the HEPA filter as the final filtration medium. Samples were taken from the inlet and outlet surfaces. Samples from the nanotextile were collected in the same manner as from the HEPA filter. QIAStat DX® 1.0 Analyzer using the Respiratory SARS CoV-2 Panel multiplex PCR detection system was selected for microorganism detection. Adenovirus was detected on the inlet surface of the HEPA filter. The outlet surface of the filter contained no viruses included in the Respiratory SARS CoV-2 Panel portfolio. The nanotextile monolayer was replaced twice during the 100h of operation, so three pieces were used and all contained coronavirus 229 E. Coronavirus 229 E was then detected in the nasopharynx of one of the members of the household as well. It may be assumed that the selected nanotextile is capable of capturing a virus of a small size.

Study on the association of Glutathione S-transferase Mu 1 (GSTM1) gene polymorphism with acute decompensated heart failure with reduced ejection fraction

The most frequent cause of hospitalization in adults over 65 in Western nations is acute decompensation of heart failure (ADHF). Due to the high death rate, it places a heavy load on healthcare systems as well as people. The present study’s objective was to ascertain the connection between polymorphisms in the Glutathione S-transferase Mu 1 (GSTM1) gene and heart-failure-reduced ejection fraction (HFrEF) patients. Sixty patients with a mean age of 25–94 were taken from both genders after the clinical diagnosis by a specialist to cases who were referred to the Ibn Al-Baitar Specialized Center for Cardiac Surgery and Ramadi Teaching Hospital, and thirty apparently healthy people were taken as a control. Blood and serum containing EDTA were drawn from sick and healthy people to extract DNA for multiplex PCR detection of the GSTM1 polymorphism. The GSTM1 genotype was found in 23 (38.33%) and the null gene in 37 (61.66%) of the 60 ADHF patients. Of 30 healthy people, 8 (26.66%) had the GSTM1 gene and 22 (73.33%) had the null gene. According to the present study, the etiology of acute decompensated heart failure (ADHF) and the antioxidant null gene (GSTM1) are related. The present study has been achieved to determine the relationship between the presence of the antioxidant gene (GSTM1) and the incidence of ADHF with reduced ejection fraction for research and therapeutic purposes, which opens new spaces in the treatment of the mentioned condition.

Direct identification of pathogens via microbial cellular DNA in whole blood by MeltArray.

Rapid identification of pathogens is critical for early and appropriate treatment of bloodstream infections. The various culture-independent assays that have been developed often have long turnaround times, low sensitivity and narrow pathogen coverage. Here, we propose a new multiplex PCR assay, MeltArray, which uses intact microbial cells as the source of genomic DNA (gDNA). The successive steps of the MeltArray assay, including selective lysis of human cells, microbial cell sedimentation, microbial cellular DNA extraction, target-specific pre-amplification and multiplex PCR detection, allowed the detection of 35 major bloodstream infectious pathogens in whole blood within 5.5 h. The limits of detection varied depending on the pathogen and ranged from 1 to 5 CFU/mL. Of 443 blood culture samples, including 373 positive blood culture samples and 70 negative blood culture samples, the MeltArray assay showed a sensitivity of 93.8% (350/373, 95% confidence interval [CI] = 90.7%-96.0%), specificity of 98.6% (69/70, 95% CI = 91.2%-99.9%), positive predictive value of 99.7% (95% CI = 98.1%-99.9%), and negative predictive value of 75.0% (95% CI = 64.7%-83.2%). The MeltArray detection results of 16 samples differed from MALDI-TOF and were confirmed by Sanger sequencing. Further testing of 110 whole blood samples from patients with suspected bloodstream infections using blood culture results revealed that the MeltArray assay had a clinical sensitivity of 100% (9/9, 95% CI = 62.8%-100.0%), clinical specificity of 74.5% (70/94, 95% CI = 64.2%-82.7%), positive predictive value of 27.3% (95% CI = 13.9%-45.8%), and negative predictive value of 100.0% (95% CI = 93.5%-100.0%). Compared with metagenomic next-generation sequencing, the MeltArray assay displayed a positive agreement of 85.7% (6/7, 95% CI = 42.0%-99.2%) and negative agreement of 100.0% (4/4, 95% CI = 39.6%-100.0%). We conclude that the MeltArray assay can be used as a rapid and reliable tool for direct identification of pathogens in bloodstream infections.

Clinical implementation of a multiplex PCR detection method for HPV causing plantar warts and genotype distribution in different geographical areas of Spain

Human papillomavirus is the ethological agent of various tumors, including plantar warts as one of the most frequent clinical presentations. Diagnosis of these warts continues to be mainly clinical, and a significant incidence of misdiagnosis leads to inadequate treatment. The aim of this study is to implement and validate a multiplex PCR detection method in the clinical setting to detect HPV in samples and to study genotype distribution in Spain to improve future molecular diagnostics. Viral DNA was extracted from 128 samples of clinically suspected plantar warts from various locations in Spain. A multiplex PCR was run alongside internal controls, and amplicons were processed for sequencing and HPV genotyping. The method was validated by assessing both inter- and intra-run repeatability. The PCR detection method returned 81.2 % (n = 104) positive results in the samples tested. Inter- and intra-run repeatability tests showed excellent intra-run agreement (κ = 1.00, p < 0.001) and good inter-run agreement (κ = 0.737, p < 0.001). The most frequent HPV type was HPV1, followed by HPV27, showing a statistical difference between the distribution of HPV genotypes in different areas of Spain. Clinical implementation of a DNA PCR detection method for plantar warts can avoid 18.8 % of unnecessary treatments in doubtful cases, and the method is reliable and validated for the purpose. HPV types show an asymmetric geographical distribution that should be considered for diagnosis and treatment.

Pathogenic characteristics of the Vibrio parahaemolyticus which caused a gastroenteritis outbreak event in Huzhou.

The pathogenic characteristics of V. parahaemolyticus isolated from a gastroenteritis outbreak event in Deqing County of Huzhou City in 2022 were analyzed. Pathogen detection was performed on 30 anal swabs (26 patients, 1 chef and 3 waiters). The isolates of V. parahaemolyticus were analyzed by serum typing, pulsed field gel electrophoresis (PFGE) molecular typing, multiplex fluorescent PCR detection of tdh/trh virulence gene and drug sensitivity test. 15 patients were positive for V. parahaemolyticus, 1 patient was positive for V. parahaemolyticus and Enteroaggregative E. coli (EAEC), 1 patient was positive for EAEC, and the chef was positive for EAEC. The serotype test results of the 16 V. parahaemolyticus were 14 O4:KUT and 2 O10:K4. All samples were negative for other tested bacteria. All V. parahaemolyticus strains were positive for tdh genes and negative for trh gene. The 16 isolates were 100% resistant to ampicillin (AMP), and sensitive to the other12 antibiotics. From the results of serotype and PFGE, the V. parahaemolyticus strains with two serotypes are clustered into two branches according to their serotypes. The three EAEC strains were non-homologous. In conclusion, we detected V. parahaemolyticus and EAEC from an outbreak of gastroenteritis. And V. parahaemolyticus with two serotypes may be the cause of this event, according to the traceability results.

Development of an ARMS multiplex real-time PCR assay for the detection of HLA-B*13:01 genotype by detecting highly specific SNPs.

HLA-B*13:01 was strongly associated with Dapsone Hypersensitivity Syndrome (DHS). This study aimed to develop and validate a rapid and economical method for HLA-B*13:01 genotyping. Two tubes multiplex real-time PCR detection system comprising amplification refractory mutation system primers and TaqMan probes was established for HLA-B*13:01 genotyping. Sequence-based typing was applied to validate the accuracy of the assay. The accuracy of the assay was 100% for HLA-B*13:01 genotyping. The detection limit of the new method was 0.025 ng DNA. The positive rate of HLA-B*13:01 in the Bouyei (20%, n = 50) populations was significantly higher than that in the Uighur population (4%, n = 100), Han (4.5%, n = 200), and Tibetan (1%, n = 100) ( P < 0.05). The proposed method is rapid and reliable for HLA-B*13:01 screening in a clinical setting.

Molecular characterization of bovine intrauterine Escherichia coli isolates in rat model

The present study was undertaken to characterize the virulence factors of Escherichia coli isolated from bovine clinical endometritis, and comparison of uterine pathology induced by different phylogroups of E. coli in a rat model. Forty E. coli isolates assigned to different phylogroups, that is, A, B1, C, D, E, F, and clad1 were used. Phenotypic detection using Congo red binding, biofilm and haemolysin production observed 77% (n=31), 55% (n=22) and 68% (n=28) positive isolates, respectively. Multiplex PCR detection of 11 VFs genes, viz; Pap, Sfa, Afa, Hly, Cnf1, Cnf2, eaeA, F41, Sta, csgA, and csgD, found only one positive isolate each for pap and csgD while 3 isolates for F41 and 8 isolates for csgA were positive. Endometritis was inducted in diestrus rats using 4 phylogroups of E. coli. Expression for COX-2 and iNOS inflammatory marker genes was observed, but there was no statistical significant difference of expression among the phylogroups. On the basis of the present findings, it may inferred that different phylogroups of E. coli exhibit different virulence characteristics, but do not differ significantly in their ability to cause uterine disease in rats.

Application of multiplex realtime PCR detection for hemorrhagic fever syndrome viruses

BackgroundMultiplex real-time PCR is a quick and cost effective method for detection of various gene simultaneously. HFSV (Hemorrhagic Fever Syndrome Virus) is a newly emerging infectious disease because of globalization and climate change. We tried to develop a molecular diagnostic technique for various causative viruses and evaluate its usefulness for improving public health. MethodsMolecular diagnostic test method that qualitatively detects viruses causing viral hemorrhagic fevers hired Taq-Man Real-time RT-PCR technique. The Ct value was experimentally observed three or more times at the RNA concentration before and after the detection limit. After designing a multiplex real-time RT-PCR test for target gene of selected 17 viruses, the detection limit for each target and the presence or absence of cross-reaction and interference reaction were evaluated to determine its availability. ResultsSix kinds of viruses, including Crimean-Congo hemorrhagic fever virus, Omsk hemorrhagic fever virus, Sabia virus, Chapare virus, Yellow fever virus, and Variola virus (A4L gene, B12R gene), were able to confirm the detection limit of 0.5 copies/μl, and other Ebola virus, Marburg virus, Rift Valley fever virus, Kyasanur Forest disease virus, Junin virus, Guanarito virus, Machupo virus, Chikungunya virus, Hantavirus, Dengue virus types 1–4, and Lassa virus (L gene, GPC gene), and 11 kinds of viruses, the detection limit was confirmed at 5 copies/μl. No cross-reaction or interference between detected genes was observed. ConclusionThe virus test method developed through this study using multiplex is expected to be used for public health and quarantine as a test method that can be used when a hemorrhagic fever virus of unknown cause is introduced.

Multiplex PCR detection of antibiotic resistance and virulence genes in multidrug-resistant Staphylococcus aureus isolated from chickens, humans, rodents, and soil in Northern Tanzania

Staphylococcus aureus (S. aureus) is a zoonotic pathogen with public health and veterinary importance. We investigated the presence of antibiotic resistance genes (ARGs) and virulence genes (VGs) in 57 multidrug-resistant (MDR) S. aureus isolated from humans (n=17), chickens (n=14), rodents (n=13), and soil (n=13) using multiplex PCR. Overall, the distribution of ARGs revealed that the tetK was found in 18/57 (31.6%), mecA in 16/57 (28.1%), tetL in 5/57 (8.9%), and ermC in 1/57 (1.8%), while ermA and tetM were not detected. For VGs, the clfB was found in 6/57 (10.5 %), coa in 8/57 (14.0%), clfA in 3/57 (5.3%), hlg in 1/57 (1.8%), ebpS in 2/57 (3.5%), fnbB in 2/57 (3.5%), luk-PV in 6/57 (10.5%) and tst in 1/57 (1.8%). Resistance genes (tetK and mecA) and virulence determinants (clfB, coa, and luk-PV) were common in all sample sources, while tst, hlg, and fnbB were specific to human, chicken, and rodent isolates, respectively. Erythromycin phenotypic resistance results correlated with the presence of ermC (r=0.42), tetL (r=0.98), and mecA (r=0.51), while tetracycline resistance correlated with tetL (r=1.00) and mecA (r=0.57) genes and methicillin resistance correlated with mecA (r=0.55) and tetL (r=0.98) genes. Positive correlations were noted between ARG (ermC) and VGs; clfA (r=0.57), hlg (r=1.00), and clfB (r=0.43), and between tetK and clfB (r=0.39); tetK and coa (r=0.36) genes. Principal component analysis (PCA) shows that tetL, ermC, and mecA contributed to tetracycline, erythromycin, and methicillin resistance, respectively. The widespread presence of resistance and virulence genes, often in combination, among MDR S. aureus in isolates from humans, chicken, rodents, and soil samples require comprehensive One-Health interventions.

Multiplex PCR detection method of genetically modified canola event (MON94100, LBFLFK, and NS-B50027-4) combined with capillary electrophoresis.

The online version contains supplementary material available at 10.1007/s10068-023-01377-z.