Table 3 Detection limit of the new established RT-RPA assay on cerebro-spinal fluid sample.

From: A recombinase polymerase amplification assay for rapid detection of rabies virus

TITERS ID50/mL

RT-qPCR

RABV-RPA

Schlottauassay

RNA molecules/reaction

Mean Cq valuea

Mean Ttb

SD

Mean Ttb

SD

Mean Ttb

SD

5.2E+05

32,033

28.42

57.68

0.480

3.10

0.020

5.33

0.030

5.2E+04

2374

31.92

61.71

0.007

4.36

0.019

7.83

0.179

5.2E+03

275

34.82

65.05

0.220

5.21

0.006

8.97

0.121

5.2E+02

82

36.45

66.92

0.672

5.45

0.086

Neg

 

5.2E+01

11

39.18

70.06

0.092

7.49

0.123

Neg

 

5.2

8

39.63

70.58

0.120

Neg

 

Neg

 

5.2E−01

 

Neg

  

Neg

 

Neg

 

5.2E−02

 

Neg

  

Neg

 

Neg

 

5.2E−03

 

Neg

  

Neg

 

Neg

 

5.2E−04

 

Neg

  

Neg

 

Neg

 

5.2E−05

 

Neg

  

Neg

 

Neg

 
  1. Tenfold serial dilutions of RABV isolate SA217695SEN with an initial virus titer of 5.2 106 ID50/mL were tested with an rabies virus RT-RPA assay previously described by Schlottau et al.21 and the new RT-RPA assay. The corresponding numbers of RNA molecules per reaction were calculated using the mean Cq values with the standard equation previously described for the RT-qPCR used here as reference technique19.
  2. Neg: negative or no fluorescence signal, Cq : quantitative cycle number, Tt: time threshold (minutes).
  3. aMean Cq value from duplicates.
  4. bMean Tt value from duplicates.
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