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Establishment and Application of Real鄄time Fluorescence Single
Primer Isothermal Amplification for
Shigella
WANG Jianchang, 摇 HU Lianxia, 摇 DUAN Yongsheng, 摇 LI Jing,摇 WANG Jinfeng
*
(
Technical Center of Inspection and Quarantine
,
Hebei Entry鄄Exit Inspection and
Quarantine Bureau
,
Shijiazhuang
050051,
China
)
Abstract
: Based on the
Shigella
ipaH gene, the RNA鄄DNA primers and Blockers were designed and syn鄄
thetized, and the real鄄time fluorescence single primer isothermal amplification ( real鄄time fluorescence
SPIA) for the detection of
Shigella
was established. The reaction time of the method was 44 min. After
detection of the 4 different
Shigella
group strains and 12 other food borne bacteria by the real鄄time fluores鄄
cent SPIA, the results showed that only the
Shigella
could be detected and showed the typical fluores鄄
cence curve. Further studies show that the sensitivity of the detection method for
Shigella flexneri
in pure
culture was 1郾 16 fg / 滋L and 1郾 3 CFU / mL. The detection limit of
Shigella flexneri
in milk was 1郾 8 CFU / mL.
The results demonstrate that the real鄄time fluorescence SPIA detection method for
Shigella
is highly sensi鄄
tive, strong specific and much more convenient.
Key words
: real鄄time fluorescence single primer isothermal amplification;
Shigella
; ipaH gene
(
责任编辑
:
叶红波
)
54
第
33
卷 第
6
期
摇 摇 摇 摇 摇 摇 摇 摇 摇
王建昌等
:
志贺氏菌实时荧光单引物等温扩增方法的建立及应用
