SYBR green-based one step quantitative real-time polymerase chain reaction assay for the detection of Zika virus in field-caught mosquitoes

Table 4 Evaluation of SYBR Green-based rRT-PCR assay performance in ZIKV infected field caught mosquitoes confirmed by a probe-based rRT-PCR assay

Sample ID^b	Probe-based rRT-PCR^a		SYBR Green-based rRT-PCR
Sample ID^b	C_q value	Result	C_q value (T_m)	Result
HT55	21.06	Positive	17.57 (80.38)	Positive
HT66	19.54	Positive	15.23 (80.24)	Positive
HT67	28.77	Positive	26.06 (80.16)	Positive
HT68	29.19	Positive	26.98 (80.22)	Positive
HT74	17.09	Positive	13.74 (80.42)	Positive
HT81	19.66	Positive	16.33 (80.1)	Positive
HT83	33.23	Positive	35 (80.04)	Positive
Ae. albopictus	Not available	Negative	Not available	Negative
Ae. albopictus	Not available	Negative	Not available	Negative
Culex spp.	Not available	Negative	Not available	Negative
Culex spp.	Not available	Negative	Not available	Negative
Ae. aegypti	Not available	Negative	Not available	Negative
Ae. aegypti	Not available	Negative	Not available	Negative
Ae. aegypti	Not available	Negative	Not available	Negative
ZIKV positive control (ATCC® VR-84)	16.77	Positive	13.84 (79.88)	Positive
Negative control	Not available	Negative	Not available	Negative

Abbreviations: C _q quantification cycle, T _m melting temperature
^aThe assay protocol has previously been described by Lanciotti et al. [9]
^bSample IDs from HT55 to HT83 are Ae. aegypti specimens collected from a zika fever cluster during the first outbreak of ZIKV in Singapore in August 2016 [6]. They were confirmed to be ZIKV-positive by the probe-based rRT-PCR assay. ZIKV negative mosquito specimens collected from the field, including the same cluster, are included for the comparison of specificity

ISSN: 1756-3305