20110201 Chipron2011 Linked In

Chipron Array R.A. Hamidjaja (Cib/LZO)

Advantage of array? ,[object Object],[object Object],Disadvantage of array? ,[object Object],[object Object]

Array format ,[object Object],[object Object],Luminex Chipron

Workflow Suspension array Planar array

Improvement (compared to Luminex) ,[object Object],[object Object],[object Object],[object Object],[object Object]

Evaluation & Validation ,[object Object],[object Object],[object Object]

Specificity B. anthracis Brucella Y. pestis C. burnetii F. tularensis B

Specificity Ixodes Anas platyrhyncos (Wild duck) Lepus europaeus (Brown hare) Y. pseudotuberculosis Homo sapiens E. coli B. cereus F. tularensis A B. thuringiensis

LOD (triplicate) Rough estimation of genomic DNA mass from 1 bacteria: 5 fg 0 0 3 3 3 2YPcaf 0 1 3 3 3 3YPpla-02 0 0 0 3 3 1YPyin Harbin 0 0 1 3 3 3YPypo Y. pestis 0 0 0 0 0 2FTpdn 0 0 0 0 0 1FTpdf-B 0 0 3 3 3 1FTwbk 3 3 3 3 3 2FTisf BD07-537 0 0 3 3 3 3FTfoa F. tularensis 0 0 0 2 3 1CBicd 0 2 3 3 3 1CBis1 0 0 3 3 3 1CBser 9 mile 0 0 3 3 3 1CBcom C. burnetii 0 0 0 0 0 1Brwbo-02 0 0 0 0 0 1Brwbo 0 1 3 3 3 1Brbcs-02 ATCC 23457 0 1 3 3 3 1Brbcs Br. melitensis 3 3 3 3 3 2BAcab 0 0 3 3 3 2BAcya-02 Vollum 493 1 0 3 3 3 1BApl3 B. anthracis 1 fg 10 fg 100 fg 1000 fg 10000 fg Probes Organism

Dynamic range Ft: F. tularensis Yp: Y. pestis

Lesson learned (1) ,[object Object],AmpXpress thermocycler AlphaHelix

SuperConvection ,[object Object],[object Object],[object Object],[object Object],[object Object],In our case: To get the same LOD and dynamic range, faster PCR is not possible Conventional thermocycler 1 X g Natural convection AmpXpress thermocycler 7000 X g Superconvection

Possible Explanations ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],Do not forget!!! Enzyme activity determines the final reaction speed

Rayleigh number and convection ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object], Ra = 0; no convection  Ra >>; the higher the rate of convection Significant contributor !!!

Epilogue ,[object Object],[object Object],[object Object],[object Object],[object Object]

Lesson learned (2) ,[object Object],[object Object],[object Object],[object Object],Cause: Biotin contamination of the capture probes Solution: If possible purchase the capture probes from other manufacturer Negative control PCR prod Water

Things to do ,[object Object],[object Object],[object Object]

20110201 Chipron2011 Linked In

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