Cross-Platform Evaluation of Commercial Real-Time SYBR Green RT-PCR Kits for Sensitive and Rapid Detection of European Bat Lyssavirus Type 1

Table 6

Comparison of the 10 commercial master mixes with regard to the practicability of the different assays and the costs, times, and number of pipetting steps.

	Ranking for cost/PCR reaction^*	Run duration (minutes)		Number of pipetting steps^**
		Rotor Gene Q MDx	Mx3005P
Two-step RT-qPCR:
SsoFast(Iscript)	9	2:42	2:53	12
QuantiTect (QuantiTect)	8	2:54	3:00	14
Maxima (Maxima)	7	2:57	3:08	14
Perfecta (qScript)	6	2:48	2:56	11
Express (SS Vilo)	10	2:50	3:01	10
One-step qRT-PCR:
Quantitect-1 step	5	2:48	2:57	7
Rotor Gene-1 step	2	2:39	2:48	7
Verso-1 step	3	2:46	2:57	8
SSIII Platinium-1 step	4	2:18	2:28	7
qScript-1 step	1	2:38	2:48	7

The cost was calculated by dividing the cost for each commercial kit, based on the manufacturer’s list price in summer of 2013 by the number of PCR reactions that could be performed with each kit. The kits were ranked from 1 to 10 based on their cost per reaction, with 1 corresponding to the least expensive and 10 to the most expensive.
Pipetting steps include those corresponding to the preparation of the master mix and the reverse transcription and PCR steps.