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Table 1 Determination of SLCMV genome copy number in SLCMV-infected sap extracts

From: Development of a triple antibody sandwich enzyme-linked immunosorbent assay for cassava mosaic disease detection using a monoclonal antibody to Sri Lankan cassava mosaic virus

Sample type

Sample name

(known copy no./5 µl)

Cq valuesb

Mean ± SD

(n = 3)

Calculated copy no./5 µl templateb

Mean ± SD

(n = 3)

Calculated copy no./ELISA wellc

Mean ± SD

(n = 3)

Plasmid standard with known copy number

STD 1 (3.00E+08)

7.19 ± 0.12

  
 

STD 2 (3.00E+07)

10.69 ± 0.15

  
 

STD 3 (3.00E+06)

14.59 ± 0.06

  
 

STD 4 (3.00E+05)

18.22 ± 0.23

  
 

STD 5 (3.00E+04)

21.42 ± 0.04

  
 

STD 6 (3.00E+03)

24.93 ± 0.09

  
 

STD 7 (3.00E+02)

28.33 ± 0.13

  
 

STD 8 (30)

31.94 ± 0.35

  

Maximum dilution of sap extract tested positive by TAS-ELISAa

Sample A dilution A7a

17.44 ± 0.22

4.08E+05 ± 5.68E+04

4.08E+06 ± 5.68E+05

 

Sample B dilution B6a

18.38 ± 0.08

2.21E+05 ± 1.19E+04

2.21E+06 ± 1.19E+05

 

Sample C dilution C5a

17.66 ± 0.23

3.55E+05 ± 5.17E+04

3.55E+06 ± 5.17E+05

  1. aThree sets of serial two-fold dilutions of SLCMV-infected plant saps were prepared from 3 infected samples and subjected to SLCMV detection by TAS-ELISA in three independent experiments. The same amount of sap extract from the maximum dilution tested positive by TAS-ELISA was used for DNA isolation and subjected to qPCR to determine the SLCMV DNA genome copy number. A7, B6 and C5 indicate the maximum dilution obtained from serial dilution sets A, B and C, respectively
  2. bData and statistical analyses were done using the CFX Manager Software version 3.1 (BIO-RAD, USA). Copy numbers were calculated from the equation y =  − 3.52x + 37.12, which was obtained from the linear regression line shown in Fig. 7
  3. cSince only 5 µl of DNA extract (1/10 of the total volume of DNA extract) were used as template for qPCR, all the results were multiplied by ten to determine the copy number of SLCMV DNA-A genomes in the maximum dilution of sap extract tested positive by TAS-ELISAa