Detection and quantification of EGFR T790M mutation in liquid biopsies by droplet digital PCR
Background: Liquid biopsy allows the identification of targetable cancer mutations in a minimally invasive manner. In patients with advanced non-small cell lung cancer (NSCLC), droplet digital PCR (ddPCR) is increasingly used to genotype the epidermal growth factor receptor (EGFR) gene in circulating cell-free DNA (cfDNA). However, the sensitivity of this method is still under debate. The aim of this study was to implement and assess the performance of a ddPCR assay for detecting the EGFR T790M mutation in liquid biopsies.
Methods: A ddPCR assay was optimized to detect the EGFR T790M mutation in plasma samples from 77 patients with NSCLC in progression.
Results: Our ddPCR assay enabled the detection and quantification of the EGFR T790M mutation at cfDNA allele frequency as low as 0.5%. The mutation was detected in 40 plasma samples, corresponding to a positivity rate of 52%.
The number of mutant molecules per mL of plasma ranged from 1 to 6,000. A re-biopsy was analyzed for 12 patients that had a negative plasma test and the mutation was detected in 2 cases. A second liquid biopsy was performed for 6 patients and the mutation was detected in 3 cases.
Conclusions: This study highlights the value of ddPCR to detect and quantify the EGFR T790M mutation in liquid biopsies in a real-world clinical setting. Our results suggest that repeated ddPCR tests in cfDNA may obviate tissue re-biopsy in patients unable to provide a tumor tissue sample suitable for molecular analysis.
A rapid and sensitive fluorescence biosensor based on plasmonic PCR
- Plasmonic PCR utilizing metallic nanoparticles has shown great advantages compared to the commercial thermocycler equipment in terms of cost, size and processing time. However, due to the strong fluorescence quenching, plasmonic nanoparticle-based PCR requires additional post-processing steps such as centrifugation and gel electrophoresis.
- This process increases the overall diagnostic time, offsetting the benefits of fast thermocycling. Here, we report a rapid and sensitive plasmonic photothermal PCR (PPT-PCR) assay method based on in situ end-point fluorescence detection.
- By using plasmonic magnetic bi-functional nanoparticles, PPT-PCR involving 30 thermocycles and fluorescence detection following magnetic separation has successfully shown that DNA targets can be detected within 5.5 minutes.
- The limit of detection (3.3 copies per μL) is comparable with that of the conventional real-time quantitative PCR; however, the assay time is about 5.5 times shorter for the PPT-PCR. The strategy of combining the photothermal effect and magnetic separation into a single particle will open new horizons in the development of fast and sensitive PCR-based biosensors for point-of care testing.
Comorbidities and Age Are Associated With Persistent COVID-19 PCR Positivity
Objectives: The impact of demographics and comorbidities on the duration of COVID-19 nasopharyngeal swab PCR positivity remains unclear. The objective of our analysis is to determine the impact of age, intensive care unit (ICU) admission, comorbidities, and ethnicity on the duration of COVID-19 PCR positivity among hospitalized patients in a large group of hospital.
Method: We studied 530 patients from a large hospital system and time to SARS-CoV-2 virus RNA PCR negativity at any-time during hospitalization or following discharge from the hospital was the primary endpoint. We included patients 18 years or older who tested positive for COVID-19 during an inpatient, outpatient, or emergency room visit between February 1, 2020, and April 14, 2020.
Results: Overall, 315 (59.4%) of our patient population continued to have a positive SARS-CoV-2 virus RNA PCR 4 weeks after the initial positive test. We found that age>70 years, chronic kidney disease, hypertension, hyperlipidemia, obesity, or coronary artery disease are associated with persistent PCR positivity for more than 4 weeks after initial diagnosis.
Conclusion: Age, and the presence of co-morbidities should be taken into consideration when interpreting a positive COVID PCR test.
Comparison of PCR and phenotypic methods for the detection of methicillin resistant Staphylococcus aureus
Background and objectives: Resistance to methicillin in methicillin resistant strains of Staphylococcus aureus (MRSA) is due to the presence of mec-A gene, which encodes a low affinity penicillin binding protein (PBP)-2a or PBP2. Accurate and rapid identification of MRSA in clinical specimens is essential for timely decision on effective treatment. The aim of the study was to compare three different methods for detection of MRSA namely cefoxitin disc diffusion, CHROM agar MRSA and VITEK-2 susceptibility with PCR which is the gold standard reference method and to find the antibiotic susceptibility pattern of these isolates by VITEK-2.
Materials and methods: A Total of 100 non-duplicate S. aureus isolates were collected from different clinical samples among both outpatient and inpatients. Detection of MRSA among these isolates was done by cefoxitin disc diffusion, VITEK-2, CHROM agar MRSA and PCR.
Results: The sensitivity and specificity of cefoxitin disc diffusion and Vitek was found to be 97.2% and 100%, while that of CHROM agar was found to be 100% and 78.6%. The overall prevalence of MRSA in our study by PCR was 72%.
Conclusion: Based on the findings in our study, isolates which show cefoxitin zone diameter < 22 mm can be reported as MRSA. However, those isolates which have a zone diameter between 22-24 mm, should ideally be confirmed by PCR.
 Alexa Fluor® 594 anti-mouse CD8a |
|||
| E16FMJ008a-100U | EnoGene | 100 μg | EUR 758.33 |
|
Description: Available in various conjugation types. |
|||
 Alexa Fluor® 594 anti-mouse CD18 |
|||
| E16FMJ018-100U | EnoGene | 100 μg | EUR 931.67 |
|
Description: Available in various conjugation types. |
|||
 Alexa Fluor® 594 anti-mouse CD19 |
|||
| E16FMJ019-100U | EnoGene | 100 μg | EUR 931.67 |
|
Description: Available in various conjugation types. |
|||
 Alexa Fluor® 594 anti-mouse CD31 |
|||
| E16FMJ031-100U | EnoGene | 100 μg | EUR 931.67 |
|
Description: Available in various conjugation types. |
|||
 Alexa Fluor® 594 anti-mouse CD45 |
|||
| E16FMJ045-100U | EnoGene | 100 μg | EUR 931.67 |
|
Description: Available in various conjugation types. |
|||
 Alexa Fluor® 594 anti-mouse CD68 |
|||
| E16FMJ068-100U | EnoGene | 100 μg | EUR 1365 |
|
Description: Available in various conjugation types. |
|||
 Alexa Fluor® 594 anti-human CD8a |
|||
| E16FHJ008a-100U | EnoGene | 100 μg | EUR 975 |
|
Description: Available in various conjugation types. |
|||
 Alexa Fluor® 594 anti-human CD14 |
|||
| E16FHJ0142-100U | EnoGene | 100 μg | EUR 975 |
|
Description: Available in various conjugation types. |
|||
 Alexa Fluor® 594 anti-human CD19 |
|||
| E16FHJ019-100U | EnoGene | 100 μg | EUR 1083.33 |
|
Description: Available in various conjugation types. |
|||
 Alexa Fluor® 594 anti-mouse CD11c |
|||
| E16FMJ011c-100U | EnoGene | 100 μg | EUR 758.33 |
|
Description: Available in various conjugation types. |
|||
 Alexa Fluor® 594 anti-mouse CD106 |
|||
| E16FMJ106-100U | EnoGene | 100 μg | EUR 931.67 |
|
Description: Available in various conjugation types. |
|||
 Alexa Fluor® 594 anti-human CD11a |
|||
| E16FHJ011a-100U | EnoGene | 100 μg | EUR 975 |
|
Description: Available in various conjugation types. |
|||
 Alexa Fluor® 594 anti-human CD11b |
|||
| E16FHJ011b-100U | EnoGene | 100 μg | EUR 931.67 |
|
Description: Available in various conjugation types. |
|||
 Mouse Anti-Human IgA - Alexa Fluor 594 |
|||
| DL87387A-100ul | DL Develop | 100 ul | EUR 190 |
|
Description: The antibody was isolated from ascitic by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Mouse Anti-Human IgA - Alexa Fluor 594 |
|||
| DL87387A-500ul | DL Develop | 500 ul | EUR 280 |
|
Description: The antibody was isolated from ascitic by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
 Mouse Anti-Human IgM - Alexa Fluor 594 |
|||
| DL87390A-100ul | DL Develop | 100 ul | EUR 155 |
|
Description: The antibody was isolated from ascitic by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Mouse Anti-Human IgM - Alexa Fluor 594 |
|||
| DL87390A-500ul | DL Develop | 500 ul | EUR 350 |
|
Description: The antibody was isolated from ascitic by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Mouse Anti-Human IgA - Alexa Fluor 594 |
|||
| E38A3634 | EnoGene | 100μL | EUR 125 |
Mouse Anti-Human IgM - Alexa Fluor 594 |
|||
| E38A3638 | EnoGene | 100μL | EUR 125 |
Mouse Anti-Human IgA - Alexa Fluor 594 |
|||
| YLF0564-100ul | Shanghai YL Biotech | 100 ul | EUR 130 |
|
Description: The antibody was isolated from ascitic by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Mouse Anti-Human IgA - Alexa Fluor 594 |
|||
| YLF0564-1ml | Shanghai YL Biotech | 1 ml | EUR 300 |
|
Description: The antibody was isolated from ascitic by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Mouse Anti-Human IgA - Alexa Fluor 594 |
|||
| YLF0564-500ul | Shanghai YL Biotech | 500 ul | EUR 210 |
|
Description: The antibody was isolated from ascitic by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Mouse Anti-Human IgM - Alexa Fluor 594 |
|||
| YLF0361-100ul | Shanghai YL Biotech | 100 ul | EUR 130 |
|
Description: The antibody was isolated from ascitic by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Mouse Anti-Human IgM - Alexa Fluor 594 |
|||
| YLF0361-1ml | Shanghai YL Biotech | 1 ml | EUR 480 |
|
Description: The antibody was isolated from ascitic by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Mouse Anti-Human IgM - Alexa Fluor 594 |
|||
| YLF0361-500ul | Shanghai YL Biotech | 500 ul | EUR 290 |
|
Description: The antibody was isolated from ascitic by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
 Rabbit Anti-Human IgM - Alexa Fluor 594 |
|||
| DL87379A-100ul | DL Develop | 100 ul | EUR 190 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Rabbit Anti-Human IgM - Alexa Fluor 594 |
|||
| DL87379A-500ul | DL Develop | 500 ul | EUR 280 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Rabbit Anti-Human IgM - Alexa Fluor 594 |
|||
| E38A3625 | EnoGene | 100μL | EUR 125 |
Rabbit Anti-Human IgM - Alexa Fluor 594 |
|||
| YLF0561-100ul | Shanghai YL Biotech | 100 ul | EUR 130 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Rabbit Anti-Human IgM - Alexa Fluor 594 |
|||
| YLF0561-1ml | Shanghai YL Biotech | 1 ml | EUR 300 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Rabbit Anti-Human IgM - Alexa Fluor 594 |
|||
| YLF0561-500ul | Shanghai YL Biotech | 500 ul | EUR 210 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
 Secondary Antibody, Alexa Fluor 594) Goat anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor 594 |
|||
| nAP-20507 | Angio Proteomie | 500ug | EUR 183.6 |
) EGFR antibody (Alexa Fluor 488) |
|||
| 61R-E109BAF | Fitzgerald | 125 ug | EUR 714 |
|
Description: Mouse monoclonal EGFR antibody (Alexa Fluor 488) |
|||
 - Alexa Fluor 594) Rabbit Anti-Bovine IgG (H&L) - Alexa Fluor 594 |
|||
| DL87366A-100ul | DL Develop | 100 ul | EUR 155 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Rabbit Anti-Bovine IgG (H&L) - Alexa Fluor 594 |
|||
| DL87366A-500ul | DL Develop | 500 ul | EUR 350 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Rabbit Anti-Bovine IgG (H&L) - Alexa Fluor 594 |
|||
| E38A3612 | EnoGene | 100μL | EUR 125 |
Rabbit Anti-Bovine IgG (H&L) - Alexa Fluor 594 |
|||
| YLF0344-100ul | Shanghai YL Biotech | 100 ul | EUR 130 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Rabbit Anti-Bovine IgG (H&L) - Alexa Fluor 594 |
|||
| YLF0344-1ml | Shanghai YL Biotech | 1 ml | EUR 480 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Rabbit Anti-Bovine IgG (H&L) - Alexa Fluor 594 |
|||
| YLF0344-500ul | Shanghai YL Biotech | 500 ul | EUR 290 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
) Alexa Fluor® 594 anti-mouse CD206 (MMR) |
|||
| E16FMJ206-100U | EnoGene | 100 μg | EUR 1191.67 |
|
Description: Available in various conjugation types. |
|||
 Goat Anti-Human IgG Fc - Alexa Fluor 594 |
|||
| DL87358A-100ul | DL Develop | 100 ul | EUR 155 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Goat Anti-Human IgG Fc - Alexa Fluor 594 |
|||
| DL87358A-500ul | DL Develop | 500 ul | EUR 350 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Goat Anti-Human IgG Fc - Alexa Fluor 594 |
|||
| YLF0339-100ul | Shanghai YL Biotech | 100 ul | EUR 130 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Goat Anti-Human IgG Fc - Alexa Fluor 594 |
|||
| YLF0339-1ml | Shanghai YL Biotech | 1 ml | EUR 480 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Goat Anti-Human IgG Fc - Alexa Fluor 594 |
|||
| YLF0339-500ul | Shanghai YL Biotech | 500 ul | EUR 290 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
 Alexa Fluor® 594 anti-human/mouse CD44 |
|||
| E16FMJ044-100U | EnoGene | 100 μg | EUR 931.67 |
|
Description: Available in various conjugation types. |
|||
 Goat Anti-Human IgG Fab - Alexa Fluor 594 |
|||
| DL87357A-100ul | DL Develop | 100 ul | EUR 190 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Goat Anti-Human IgG Fab - Alexa Fluor 594 |
|||
| DL87357A-500ul | DL Develop | 500 ul | EUR 280 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
 Mouse Anti-Human IgG Fc - Alexa Fluor 594 |
|||
| DL87389A-100ul | DL Develop | 100 ul | EUR 190 |
|
Description: The antibody was isolated from ascitic by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Mouse Anti-Human IgG Fc - Alexa Fluor 594 |
|||
| DL87389A-500ul | DL Develop | 500 ul | EUR 280 |
|
Description: The antibody was isolated from ascitic by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Goat Anti-Human IgG Fab - Alexa Fluor 594 |
|||
| E38A3603 | EnoGene | 100μL | EUR 125 |
Mouse Anti-Human IgG Fc - Alexa Fluor 594 |
|||
| E38A3636 | EnoGene | 100μL | EUR 125 |
Goat Anti-Human IgG Fab - Alexa Fluor 594 |
|||
| YLF0558-100ul | Shanghai YL Biotech | 100 ul | EUR 130 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Goat Anti-Human IgG Fab - Alexa Fluor 594 |
|||
| YLF0558-1ml | Shanghai YL Biotech | 1 ml | EUR 300 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Goat Anti-Human IgG Fab - Alexa Fluor 594 |
|||
| YLF0558-500ul | Shanghai YL Biotech | 500 ul | EUR 210 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Mouse Anti-Human IgG Fc - Alexa Fluor 594 |
|||
| YLF0566-100ul | Shanghai YL Biotech | 100 ul | EUR 130 |
|
Description: The antibody was isolated from ascitic by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Mouse Anti-Human IgG Fc - Alexa Fluor 594 |
|||
| YLF0566-1ml | Shanghai YL Biotech | 1 ml | EUR 300 |
|
Description: The antibody was isolated from ascitic by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Mouse Anti-Human IgG Fc - Alexa Fluor 594 |
|||
| YLF0566-500ul | Shanghai YL Biotech | 500 ul | EUR 210 |
|
Description: The antibody was isolated from ascitic by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
 Alexa Fluor® 594 anti-human/mouse CD11b |
|||
| E16FMJ011b-100U | EnoGene | 100 μg | EUR 931.67 |
|
Description: Available in various conjugation types. |
|||
 Rabbit Anti-Human IgG Fc - Alexa Fluor 594 |
|||
| DL87376A-100ul | DL Develop | 100 ul | EUR 155 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Rabbit Anti-Human IgG Fc - Alexa Fluor 594 |
|||
| DL87376A-500ul | DL Develop | 500 ul | EUR 350 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
 Mouse Anti-Human IgG Fab - Alexa Fluor 594 |
|||
| DL87388A-100ul | DL Develop | 100 ul | EUR 190 |
|
Description: The antibody was isolated from ascitic by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Mouse Anti-Human IgG Fab - Alexa Fluor 594 |
|||
| DL87388A-500ul | DL Develop | 500 ul | EUR 280 |
|
Description: The antibody was isolated from ascitic by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Rabbit Anti-Human IgG Fc - Alexa Fluor 594 |
|||
| E38A3622 | EnoGene | 100μL | EUR 125 |
Mouse Anti-Human IgG Fab - Alexa Fluor 594 |
|||
| E38A3635 | EnoGene | 100μL | EUR 125 |
Mouse Anti-Human IgG Fab - Alexa Fluor 594 |
|||
| YLF0565-100ul | Shanghai YL Biotech | 100 ul | EUR 130 |
|
Description: The antibody was isolated from ascitic by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Mouse Anti-Human IgG Fab - Alexa Fluor 594 |
|||
| YLF0565-1ml | Shanghai YL Biotech | 1 ml | EUR 300 |
|
Description: The antibody was isolated from ascitic by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Mouse Anti-Human IgG Fab - Alexa Fluor 594 |
|||
| YLF0565-500ul | Shanghai YL Biotech | 500 ul | EUR 210 |
|
Description: The antibody was isolated from ascitic by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Rabbit Anti-Human IgG Fc - Alexa Fluor 594 |
|||
| YLF0354-100ul | Shanghai YL Biotech | 100 ul | EUR 130 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Rabbit Anti-Human IgG Fc - Alexa Fluor 594 |
|||
| YLF0354-1ml | Shanghai YL Biotech | 1 ml | EUR 480 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Rabbit Anti-Human IgG Fc - Alexa Fluor 594 |
|||
| YLF0354-500ul | Shanghai YL Biotech | 500 ul | EUR 290 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
 (Alexa Fluor 594)) Donkey anti Rat IgG (H + L) (Alexa Fluor 594) |
|||
| 43R-ID022AF | Fitzgerald | 500 ug | EUR 334 |
|
Description: Donkey anti Rat IgG (H + L) secondary antibody (Alexa Fluor 594) |
|||
Donkey anti Rat IgG (H + L) (Alexa Fluor 594) |
|||
| 43R-ID047AF | Fitzgerald | 500 ug | EUR 465 |
|
Description: Donkey anti Rat IgG (H + L) secondary antibody (Alexa Fluor 594) |
|||
 - Alexa Fluor 594) Goat Anti-Pig IgG (H&L) - Alexa Fluor 594 |
|||
| DL87364A-100ul | DL Develop | 100 ul | EUR 155 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Goat Anti-Pig IgG (H&L) - Alexa Fluor 594 |
|||
| DL87364A-500ul | DL Develop | 500 ul | EUR 350 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Goat Anti-Pig IgG (H&L) - Alexa Fluor 594 |
|||
| E38A3610 | EnoGene | 100μL | EUR 125 |
Goat Anti-Pig IgG (H&L) - Alexa Fluor 594 |
|||
| YLF0343-100ul | Shanghai YL Biotech | 100 ul | EUR 130 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Goat Anti-Pig IgG (H&L) - Alexa Fluor 594 |
|||
| YLF0343-1ml | Shanghai YL Biotech | 1 ml | EUR 480 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Goat Anti-Pig IgG (H&L) - Alexa Fluor 594 |
|||
| YLF0343-500ul | Shanghai YL Biotech | 500 ul | EUR 290 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
 (Alexa Fluor 594)) Donkey anti Goat IgG (H + L) (Alexa Fluor 594) |
|||
| 43R-ID005AF | Fitzgerald | 500 ug | EUR 333 |
|
Description: Donkey anti Goat IgG (H + L) secondary antibody (Alexa Fluor 594) |
|||
 - Alexa Fluor 594) Mouse Anti-Pig IgG (H&L) - Alexa Fluor 594 |
|||
| DL87393A-100ul | DL Develop | 100 ul | EUR 190 |
|
Description: The antibody was isolated from ascitic by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Mouse Anti-Pig IgG (H&L) - Alexa Fluor 594 |
|||
| DL87393A-500ul | DL Develop | 500 ul | EUR 280 |
|
Description: The antibody was isolated from ascitic by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Mouse Anti-Pig IgG (H&L) - Alexa Fluor 594 |
|||
| E38A3641 | EnoGene | 100μL | EUR 125 |
Mouse Anti-Pig IgG (H&L) - Alexa Fluor 594 |
|||
| YLF0569-100ul | Shanghai YL Biotech | 100 ul | EUR 130 |
|
Description: The antibody was isolated from ascitic by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Mouse Anti-Pig IgG (H&L) - Alexa Fluor 594 |
|||
| YLF0569-1ml | Shanghai YL Biotech | 1 ml | EUR 300 |
|
Description: The antibody was isolated from ascitic by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Mouse Anti-Pig IgG (H&L) - Alexa Fluor 594 |
|||
| YLF0569-500ul | Shanghai YL Biotech | 500 ul | EUR 210 |
|
Description: The antibody was isolated from ascitic by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
 - Alexa Fluor 594) Goat Anti-Human IgG (H&L) - Alexa Fluor 594 |
|||
| DL87356A-100ul | DL Develop | 100 ul | EUR 140 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Goat Anti-Human IgG (H&L) - Alexa Fluor 594 |
|||
| DL87356A-500ul | DL Develop | 500 ul | EUR 330 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
 - Alexa Fluor 594) Goat Anti-Mouse IgG (H&L) - Alexa Fluor 594 |
|||
| DL87362A-100ul | DL Develop | 100 ul | EUR 140 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Goat Anti-Mouse IgG (H&L) - Alexa Fluor 594 |
|||
| DL87362A-500ul | DL Develop | 500 ul | EUR 330 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
 - Alexa Fluor 594) Rabbit Anti-Cat IgG (H&L) - Alexa Fluor 594 |
|||
| DL87367A-100ul | DL Develop | 100 ul | EUR 155 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Rabbit Anti-Cat IgG (H&L) - Alexa Fluor 594 |
|||
| DL87367A-500ul | DL Develop | 500 ul | EUR 350 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
 - Alexa Fluor 594) Rabbit Anti-Dog IgG (H&L) - Alexa Fluor 594 |
|||
| DL87369A-100ul | DL Develop | 100 ul | EUR 155 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Rabbit Anti-Dog IgG (H&L) - Alexa Fluor 594 |
|||
| DL87369A-500ul | DL Develop | 500 ul | EUR 350 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
 - Alexa Fluor 594) Rabbit Anti-Pig IgG (H&L) - Alexa Fluor 594 |
|||
| DL87382A-100ul | DL Develop | 100 ul | EUR 155 |
|
Description: The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. |
|||
Universal primers for rapid detection of six pospiviroids in Solanaceae plants using one-step RT-PCR and RT-LAMP
A number of viruses and viroids infect solanaceous plants causing severe yield losses. Several seed-borne viroids are currently listed as quarantine pathogens in many countries. Among them, columnea latent viroid (CLVd), pepper chat fruit viroid (PCFVd), potato spindle tuber viroid (PSTVd), tomato apical stunt viroid (TASVd), tomato chlorotic dwarf viroid (TCDVd), and tomato planta macho viroid (TPMVd) are of major concerns.
The objective of this study was to design and test universal primers that could be used to detect six viroids in solanaceous plants using one-step RT-PCR and reverse transcription loop-mediated isothermal amplification (RT-LAMP). Results revealed that a pair of degenerate primers could be used in a one-step RT-PCR to amplify six pospiviroids from Solanaceae seeds and plants. Moreover, five primers were designed and used in RT-LAMP to amplify six pospiviroids.
The minimal concentration of viroid RNA required for a successful detection varied, ranging from one femtogram to 10 nanograms, depending on the species of viroid and detection method. In general, RT-LAMP was more sensitive than RT-PCR but both assays were rapid and highly sensitive tools to detect six pospiviroids.
Detection methods currently in use for these viroids require at least two different sets of primers. The assays developed in this research could facilitate to screen a large number of solanaceous plants and seeds intended for import and export.