• juni 30, 2021
sexpert-vlaanderen

Klinisch nut van Droplet Digital PCR om BCR-ABL1

Klinisch nut van Droplet Digital PCR om BCR-ABL1-transcripten van patiënten met Philadelphia-chromosoom-positieve acute lymfoblastische leukemie te monitoren Publish-chimere antigeenreceptor 19/22 T-celcocktailtherapie

Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph+ ALL) accounts for 20-30% of grownup sufferers with ALL, characterised by translocation of t (9, 22). Tyrosine kinase inhibitors (TKIs) have considerably improved the result despite the fact that there are nonetheless some issues together with relapse attributable to drug-resistant mutations and suboptimal molecular remission depth. Beforehand, we reported the security and efficacy of sequential infusion of CD19/22 chimeric antigen receptor T-cell (CAR-T) immunotherapy within the remedy of relapsed/refractory (R/R) B-cell neoplasms together with instances with Ph+ ALL. Given doable deeper response, extra sufferers had been anticipated to achieve optimum minimal residual illness (MRD) response. Another technique, duplex droplet digital PCR (ddPCR) with excessive sensitivity was established, which may present absolute quantification of MRD with out the necessity for calibration curves. Right here, we retrospectively collected 95 bone marrow samples from 10 sufferers with R/R Ph+, who acquired 19/22 CAR-T-cell cocktail remedy.

Notably, sequential molecular remission for greater than three months (SMR3), a major indicator based mostly on ddPCR after CAR-T infusion was established, which was outlined as a sequential molecular remission for not <three months with destructive MRD. On this cohort, no recurrence was noticed in six sufferers reaching SMR3, the place 4 of whom accepted allogeneic hematopoietic stem cell transplantation (allo-HSCT) after CAR-T cell routine. Sadly, the opposite 4 sufferers who didn’t attain SMR3 relapsed, and didn’t obtain additional particular remedy besides CAR-T routine. To sum up, ddPCR could also be an alternate, particularly when nucleic acid was inadequate in scientific apply. No achievement of SMR3 could also be an early warning of potential relapse after CAR-T and indicating the initiation of different therapies together with allo-HSCT.

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2×Taq PCR Master Mix(with dye)

K1034-50 50×1 ml
EUR 616.8

2 × Taq Plus Master Mix

P211-01 5 ml
EUR 160.8

2 × Taq Plus Master Mix

P211-02 15 ml
EUR 231.6

2 × Taq Plus Master Mix

P211-03 50 ml
EUR 466.8

2 × Rapid Taq Master Mix

P222-01 5 ml (5×1ml)
EUR 144

2 × Rapid Taq Master Mix

P222-02 15 ml (15×1ml)
EUR 183.6

2 × Rapid Taq Master Mix

P222-03 50 ml (50 x 1 ml)
EUR 308.4

2 × Rapid Taq Master Mix

P222-04 50 ml (10×5ml)
EUR 308.4

Accuris Taq Plus Master Mix

PR1001-TP-1000 1 PC
EUR 992.89

Accuris Taq Plus Master Mix

PR1001-TP-200 1 PC
EUR 291.41

Accuris Taq Plus Master Mix

PR1001-TP-S 1 PC
EUR 83.75

2× EpiArt HS Taq Master Mix

EM201-01 1ml
EUR 174

2× EpiArt HS Taq Master Mix

EM201-02 5 ml (5×1ml)
EUR 363.6

2× EpiArt HS Taq Master Mix

EM201-03 15 ml (15×1ml)
EUR 770.4

2 × Taq Master Mix (Dye Plus)

P112-01 5 ml
EUR 140.4

2 × Taq Master Mix (Dye Plus)

P112-02 15 ml
EUR 175.2

2 × Taq Master Mix (Dye Plus)

P112-03 50 ml
EUR 283.2

Accuris Hot Start Taq Master Mix

PR1001-HS-1000 1 PC
EUR 589.04

Accuris Hot Start Taq Master Mix

PR1001-HS-200 1 PC
EUR 203.36

Accuris Taq Master Mix Red Dye

PR1001-R-1000 1 PC
EUR 360.31

Accuris Taq Master Mix Red Dye

PR1001-R-200 1 PC
EUR 145.94

Accuris Taq Master Mix Red Dye

PR1001-R-S 1 PC
EUR 83.75

Taq PCR Master Mix (2X, Blue Dye)

BS9295 1ml
EUR 101.76

Taq PCR Master Mix (2X, Blue Dye)

BS9296 5ml
EUR 227.04

Taq PCR Master Mix (2X, Red Dye)

BS9297 1ml
EUR 101.76

Taq PCR Master Mix (2X, Red Dye)

BS9298 5ml
EUR 227.04

2 × Taq Plus Master Mix (Dye Plus)

P212-01 5 ml
EUR 159.6

2 × Taq Plus Master Mix (Dye Plus)

P212-02 15 ml
EUR 231.6

2 × Taq Plus Master Mix (Dye Plus)

P212-03 50 ml
EUR 466.8

2× EpiArt HS Taq Master Mix (Dye Plus)

EM202-01 1ml
EUR 174

2× EpiArt HS Taq Master Mix (Dye Plus)

EM202-02 5 ml (5×1ml)
EUR 363.6

2× EpiArt HS Taq Master Mix (Dye Plus)

EM202-03 15 ml (15×1ml)
EUR 770.4

2 × Taq Plus Master Mix II (Dye Plus)

P213-01 5 ml
EUR 160.8

2 × Taq Plus Master Mix II (Dye Plus)

P213-02 15 ml
EUR 231.6

2 × Taq Plus Master Mix II (Dye Plus)

P213-03 50 ml
EUR 466.8

Accuris Hot Start Taq Master Mix Red Dye

PR1001-HSR-1000 1 PC
EUR 589.04

Accuris Hot Start Taq Master Mix Red Dye

PR1001-HSR-200 1 PC
EUR 203.36

Accuris Hot Start Taq Master Mix Red Dye

PR1001-HSR-S 1 PC
EUR 83.75

Taq DNA Polymerase with dNTP Mix (6000U)

9K-001-0018 6000U
EUR 1536.74

Taq DNA Polymerase with dNTP Mix (500U)

9K-001-0031 500U
EUR 351.28

Taq DNA Polymerase with dNTP Mix (3000U)

9K-001-0032 3000U
EUR 1088.87

Taq DNA Polymerase with dNTP Mix (1000U)

9K-001-0034 1000U
EUR 518.84

Green Taq Mix

P131-01 5 ml
EUR 142.8

Green Taq Mix

P131-02 15 ml
EUR 180

Green Taq Mix

P131-03 50 ml
EUR 297.6

Jade? Master Mix

M1105-500 each
EUR 529.2

Taqman Master Mix

M1121-500 each
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Fast Probe Master Mix with Rox (200 rxn)

31016 2x1mL
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Description: Minimum order quantity: 1 unit of 2x1mL

Fast Probe Master Mix with Rox (500 rxn)

31016-1 5x1mL
EUR 559.2
Description: Minimum order quantity: 1 unit of 5x1mL

Fast Probe Master Mix with Rox (5000 rxn)

31016-2 50x1mL
EUR 4645.2
Description: Minimum order quantity: 1 unit of 50x1mL

Taq DNA Polymerase - 5,000 units with separate dNTP Mix

3245d 1/EA
EUR 802.8

Fast-Taq DNA Polymerase with 10mM dNTP Mix (500U)

9K-001-0003 500U
EUR 575.22

Fast-Taq DNA Polymerase with 10mM dNTP Mix (2500U)

9K-001-0004 2500U
EUR 2465.38

High-Taq DNA Polymerase with 10mM dNTP Mix (250U)

9K-001-0005 250U
EUR 384.17

High-Taq DNA Polymerase with 10mM dNTP Mix (4x250U)

9K-001-0006 4x250U
EUR 1055.98

High-Taq DNA Polymerase with 10mM dNTP Mix (5000U)

9K-001-0020 5000U
EUR 2878.8

amfiSure PCR Master Mix

P0311-010 2x50 rxns
EUR 151.2

amfiSure PCR Master Mix

P0311-025 5X50 rxns
EUR 198

amfiSure PCR Master Mix

P0311-050 10X50 rxns
EUR 262.8

amfiSure PCR Master Mix

P0311-100 10x1ml
EUR 343.2

amfiSure PCR Master Mix

P0311-125 15X50 rxns
EUR 333.6

amfiSure PCR Master Mix

P0311-200 20x1ml
EUR 562.8

amfiSure PCR Master Mix

P0311-250 50x50 rxns
EUR 902.4

amfiSure PCR Master Mix

P0311-500 100x50 rxns
EUR 1509.6

amfiXpand PCR Master Mix

P0331-010 2X50 rxns
EUR 181.2

amfiXpand PCR Master Mix

P0331-025 5X50 rxns
EUR 327.6

amfiXpand PCR Master Mix

P0331-050 10X50 rxns
EUR 537.6

amfiXpand PCR Master Mix

P0331-250 50x50 rxns
EUR 2247.6

2 × AceTaq Master Mix

P411-01 1 ml
EUR 142.8

2 × AceTaq Master Mix

P411-02 5 ml
EUR 189.6

2 × AceTaq Master Mix

P411-03 15 ml
EUR 314.4

Taqman Master Mix-iCycler

M1122-500 each
EUR 529.2

Taqman Master Mix-ROX

M1124-500 each
EUR 529.2

Taqman Master Mix-Multiplex

M1125-500 each
EUR 529.2

Volatile Organics Mix

60-BIG-MIX 1ML
EUR 120.84

5 Component Mix

AOAC-MIX-1 EACH
EUR 117.42

Seamless cloning Master Mix (Kit)

B632219 40RXN, 40prep
EUR 319.96

amfiSure Prime PCR Master Mix

P1311-025 5X50 rxns
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amfiSure Prime PCR Master Mix

P1311-050 10X50 rxns
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amfiSure Prime PCR Master Mix

P1311-125 15X50 rxns
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amfiSure Prime PCR Master Mix

P1311-500 100x50 rxns
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SYBR Green qPCR Master Mix

HY-K0501 5 mL (500 rxns)
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RT Master Mix for qPCR

HY-K0510 1 mL (100 rxns)
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amfiSure Advanced PCR Master Mix

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amfiSure Advanced PCR Master Mix

P2311-050 10X50 rxns
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amfiSure Advanced PCR Master Mix

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amfiSure Advanced PCR Master Mix

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amfiSure Advanced PCR Master Mix

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2 × Vazyme LAmp Master Mix

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2 × Vazyme LAmp Master Mix

P311-02 5 ml
EUR 254.4

Kwantitatieve detectie en monitoring van Colletotrichum siamense in rubberbomen met behulp van real-time PCR

Colletotrichum siamense is among the most vital pathogens of rubber bushes in Asia. The correct detection and quantification of C. siamense populations in rubber bushes are of significance for monitoring the epidemics of the illness. On this research, we developed an ITS-based real-time PCR technique to effectively detect C. siamense infecting rubber tree, which reliably detected as little as 100 fg genomic DNA, 100 copies of goal DNA and 20 conidia. The actual-time PCR protocol acknowledged all C. siamense isolates collected from three provinces in China, whereas no amplification was noticed with rubber tree and its different pathogens. Detection and quantification of C. siamense had been carried out in artificially and naturally contaminated rubber leaves.

We may nonetheless detect C. siamense in plant mixes of which solely 0.0001% of the tissue contaminated. An accumulation of C. siamense DNA was noticed throughout the entire an infection course of in any respect three leaf phenological levels, suggesting the real-time PCR technique can be utilized to observe C. siamense improvement in rubber bushes. Lastly, the strategy allowed the detection of C. siamense in naturally contaminated and symptomless leaves of rubber bushes within the fields. In contrast with earlier detection strategies, the real-time PCR technique is extra particular and extra delicate, and will likely be of nice use for research aiming to realize a greater understanding of the epidemiology of Colletotrichum leaf illness, in addition to the prediction of illness threat and the management proposal.

Integratie van microfluïdische monstervoorbereiding met PCR-detectie om de effecten van gelijktijdige scheiding van DNA-remmers en uitwisseling van DNA-oplossingen te onderzoeken

On this paper, we utilized a curved-channel microfluidic machine to separate DNA from PCR-inhibitor-containing water and concurrently wash them into clear water for detection utilizing a conveyable PCR thermocycler. Environmental DNA (eDNA) sampling has turn into an efficient surveying strategy for detecting uncommon organisms. Nevertheless, low focus eDNA molecules could also be masked by PCR inhibitors throughout amplification and detection, rising the chance of false negatives. Due to this fact, applied sciences for on-site DNA separation and washing are urgently wanted. Our machine consisted of a half-circle microchannel with a DNA-inhibitor pattern inlet, a clear buffer inlet, and a number of retailers.

By utilizing the flow-induced inertial forces, 10 μm DNA-conjugated microparticles had been centered on the inner-wall of the curved microchannel whereas separation from 1 μm inhibitor-conjugated microparticles and DNA washing had been achieved concurrently with the Dean circulate. We achieved singleplex focusing, isolation and washing of 10 μm particles at an effectivity of 94.5 ± 2.0%. In duplex experiments with 1 μm and 10 μm particles, bigger particles had been washed with an effectivity of 92.1 ± 1.6% and a purity of 79 ± 2%.

By surface-functionalizing the microparticles with affinity teams in opposition to Atlantic salmon DNA and humic acid (HA), and processing samples of varied concentrations in our machine, we achieved an efficient purification and detection of DNA molecules utilizing the transportable PCR thermocycler. Our technique considerably decreased PCR quantitation cycles from Cq > 38 to Cq = 30.35 ± 0.5, which confirmed enhancement of PCR amplification. The proposed machine takes a promising step ahead in pattern preparation in the direction of an built-in machine that can be utilized for simultaneous purification and answer alternate of DNA in point-of-need environmental monitoring purposes.

SARS-CoV-2-serologie verhoogt diagnostische nauwkeurigheid bij CT-vermoede, PCR-negatieve COVID-19-patiënten tijdens pandemie 

Background: Within the absence of PCR detection of SARS-CoV-2 RNA, correct prognosis of COVID-19 is difficult. Low-dose computed tomography (CT) detects pulmonary infiltrates with excessive sensitivity, however findings could also be non-specific. This research assesses the diagnostic worth of SARS-CoV-2 serology for sufferers with distinct CT options however destructive PCR.

Strategies: IgM/IgG chemiluminescent immunoassay was carried out for 107 sufferers with confirmed (group A: PCR + ; CT ±) and 46 sufferers with suspected (group B: repetitive PCR-; CT +) COVID-19, admitted to a German college hospital through the pandemic’s first wave. A standardized, in-house CT classification of radiological indicators of a viral pneumonia was used to evaluate the likelihood of COVID-19.

Outcomes: Seroconversion charges (SR) decided on day 5, 10, 15, 20 and 25 after symptom onset (SO) had been 8%, 25%, 65%, 76% and 91% for group A, and 0%, 10%, 19%, 37% and 46% for group B, respectively; (p < 0.01). In comparison with hospitalized sufferers with a non-complicated course (non-ICU sufferers), seroconversion tended to happen at decrease frequency and delayed in sufferers on intensive care items. SR of sufferers with CT findings categorised as excessive certainty for COVID-19 had been 8%, 22%, 68%, 79% and 93% in group A, in contrast with 0%, 15%, 28%, 50% and 50% in group B (p < 0.01). SARS-CoV-2 serology established a particular prognosis in 12/46 group B sufferers. In 88% (8/9) of sufferers with destructive serology > 14 days after symptom onset (group B), clinico-radiological consensus reassessment revealed possible diagnoses apart from COVID-19. Sensitivity of SARS-CoV-2 serology was superior to PCR > 17d after symptom onset.

Conclusions: Roughly one-third of sufferers with distinct COVID-19 CT findings are examined destructive for SARS-CoV-2 RNA by PCR rendering appropriate prognosis tough. Implementation of SARS-CoV-2 serology testing alongside present CT/PCR-based diagnostic algorithms improves discrimination between COVID-19-related and non-related pulmonary infiltrates in PCR destructive sufferers. Nevertheless, sensitivity of SARS-CoV-2 serology strongly will depend on the time of testing and turns into superior to PCR after the twond week following symptom onset.

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