Pseudomonas aeruginosa
(
P. aeruginosa
) is a common opportunistic
Gram-negative
pathogen that may cause infections to immunocompromised patients.
However, sensitive and reliable analysis of
P. aeruginosa
remains a huge challenge. In this method, target recognition assists
the formation of a self-primer and initiates single-stranded chain
production. The produced single-stranded DNA chain is identified by
CRISPR-Cas12a, and consequently, the
trans
-cleavage
activity of the Cas12a enzyme is activated to parallelly digest Ag
+
aptamer sequences that are chelated with silver ions (Ag
+
). The released Ag
+
reacted with 3,3′,5,5′-tetramethylbenzidine
(TMB) for coloring. Compared with the traditional color developing
strategies, which mainly rely on the DNA hybridization, the color
developing strategy in this approach exhibits a higher efficiency
due to the robust
trans
-cleavage activity of the
Cas12a enzyme. Consequently, the method shows a low limit of detection
of a wide detection of 5 orders of magnitudes and a low limit of detection
of 21 cfu/mL, holding a promising prospect in early diagnosis of infections.
Herein, we develop a sensitive and reliable method for direct and
colorimetric detection of
P. aeruginosa
by integrating self-primer-assisted chain production and CRISPR-Cas12a-based
color reaction and believe that the established approach will facilitate
the development of bacteria-analyzing sensors.