Visual and Rapid Detection of Acinetobacter Baumannii by a Multiple Cross Displacement Amplification Combined with Nanoparticles-based Biosensor Assay

Overview

Journal AMB Express

Date 2019 Feb 27

PMID 30806854

Citations 7

Authors

Xueqin Cheng

Jing Yang

Meifang Wang

Peng Wu

Qiong Du

Jinjuan He

Yijun Tang

Affiliations

Soon will be listed here.

Abstract

The traditional microbiological methods used for detecting Acinetobacter baumannii were usually time-consuming and labor-intensive. Thus, we sought to establish a novel rapid detecting method for target pathogen. A set of multiple cross displacement amplification (MCDA) primers was designed to recognize 10 different regions of the pgaD gene, which was conservative and specific for the bacterium. In the MCDA system, amplification primers D1 and R1 were 5'-labeled with FITC (fluorescein) and biotin, respectively. Numerous FITC- and biotin-attached duplex amplicons were formed during the amplification stage, which were detected by nanoparticles-based lateral flow biosensors (LFB) through immunoreactions (FITC on the duplex and anti-FITC on the LFB test line) and biotin/streptavidin interaction (biotin on the duplex and streptavidin on the nanoparticles). The results showed that the optimized reaction condition of MCDA-LFB method was 62 °C within 25 min. There was no cross reaction with non-A. baumannii species and the non-Acinetobacter genera, and the detection limit for DNA samples was 100 fg/reaction. For 135 sputum samples, the detection results showed that the detection ability of MCDA-LFB assay was superior to the culture methods and conventional PCR. Therefore, MCDA-LFB assay could be a potential tool for the rapid detection of A. baumannii in clinical samples and low resource areas.

Citing Articles

Treatment and Management of Pneumonia: Lessons Learned from Recent World Event.

Rangel K, De-Simone S Infect Drug Resist. 2024; 17:507-529.

PMID: 38348231 PMC: 10860873. DOI: 10.2147/IDR.S431525.

Rapid and Visual Detection of SARS-CoV-2 Using Multiplex Reverse Transcription Loop-Mediated Isothermal Amplification Linked With Gold Nanoparticle-Based Lateral Flow Biosensor.

Chen X, Zhou Q, Li S, Yan H, Chang B, Wang Y Front Cell Infect Microbiol. 2021; 11:581239.

PMID: 34336708 PMC: 8316814. DOI: 10.3389/fcimb.2021.581239.

Development and Application of a Multiple Cross Displacement Amplification Combined With Nanoparticle-Based Lateral Flow Biosensor Assay to Detect .

Wang Y, Zhao X, Cheng J, Tang X, Chen X, Yu H Front Microbiol. 2021; 12:681488.

PMID: 34177867 PMC: 8222920. DOI: 10.3389/fmicb.2021.681488.

Highly specific and sensitive detection of the Mycobacterium tuberculosis complex using multiplex loop-mediated isothermal amplification combined with a nanoparticle-based lateral flow biosensor.

Chen X, Huang J, Xiao Z, Yang X, Chen Y, Zheng W Braz J Microbiol. 2021; 52(3):1315-1325.

PMID: 34176103 PMC: 8324766. DOI: 10.1007/s42770-021-00520-4.

Highly Sensitive and Rapid Identification of Based on Multiple Cross Displacement Amplification Coupled With Lateral Flow Biosensor Assay.

Cheng X, Dou Z, Yang J, Gu Y, Liu D, Xie L Front Microbiol. 2020; 11:1926.

PMID: 32983004 PMC: 7485445. DOI: 10.3389/fmicb.2020.01926.

References

Van Looveren M, Goossens H . Antimicrobial resistance of Acinetobacter spp. in Europe. Clin Microbiol Infect. 2004; 10(8):684-704. DOI: 10.1111/j.1469-0691.2004.00942.x. View

Higgins P, Wisplinghoff H, Krut O, Seifert H . A PCR-based method to differentiate between Acinetobacter baumannii and Acinetobacter genomic species 13TU. Clin Microbiol Infect. 2007; 13(12):1199-201. DOI: 10.1111/j.1469-0691.2007.01819.x. View

Dijkshoorn L, Nemec A, Seifert H . An increasing threat in hospitals: multidrug-resistant Acinetobacter baumannii. Nat Rev Microbiol. 2007; 5(12):939-51. DOI: 10.1038/nrmicro1789. View

Choi A, Slamti L, Avci F, Pier G, Maira-Litran T . The pgaABCD locus of Acinetobacter baumannii encodes the production of poly-beta-1-6-N-acetylglucosamine, which is critical for biofilm formation. J Bacteriol. 2009; 191(19):5953-63. PMC: 2747904. DOI: 10.1128/JB.00647-09. View

Marioni G, Marchese-Ragona R, Boldrin C, Parisi S, Vianello A, Prencipe R . Deep neck abscess due to Acinetobacter baumannii infection. Am J Otolaryngol. 2009; 31(4):304-7. DOI: 10.1016/j.amjoto.2009.02.018. View

Turton J, Shah J, Ozongwu C, Pike R . Incidence of Acinetobacter species other than A. baumannii among clinical isolates of Acinetobacter: evidence for emerging species. J Clin Microbiol. 2010; 48(4):1445-9. PMC: 2849580. DOI: 10.1128/JCM.02467-09. View

Chiang M, Kuo S, Chen Y, Lee Y, Chen T, Fung C . Polymerase chain reaction assay for the detection of Acinetobacter baumannii in endotracheal aspirates from patients in the intensive care unit. J Microbiol Immunol Infect. 2011; 44(2):106-10. DOI: 10.1016/j.jmii.2010.04.003. View

Durante-Mangoni E, Zarrilli R . Global spread of drug-resistant Acinetobacter baumannii: molecular epidemiology and management of antimicrobial resistance. Future Microbiol. 2011; 6(4):407-22. DOI: 10.2217/fmb.11.23. View

Zhang L, Ding G, Wei L, Pan X, Mei L, Zhang Y . Establishment of a novel target-based real-time quantitative PCR method for Acinetobacter baumannii detection. Diagn Mol Pathol. 2011; 20(4):242-8. DOI: 10.1097/PDM.0b013e31821bbb1e. View

10.

McConnell M, Perez-Ordonez A, Perez-Romero P, Valencia R, Lepe J, Vazquez-Barba I . Quantitative real-time PCR for detection of Acinetobacter baumannii colonization in the hospital environment. J Clin Microbiol. 2012; 50(4):1412-4. PMC: 3318517. DOI: 10.1128/JCM.06566-11. View

11.

Nomanpour B, Ghodousi A, Babaei A, Abtahi H, Tabrizi M, Feizabadi M . Rapid, cost-effective, sensitive and quantitative detection of Acinetobacter baumannii from pneumonia patients. Iran J Microbiol. 2012; 3(4):162-9. PMC: 3330178. View

12.

Rice L, Reis Jr A, Ronish B, Carver-Brown R, Czajka J, Gentile N . Design of a single-tube, endpoint, linear-after-the-exponential-PCR assay for 17 pathogens associated with sepsis. J Appl Microbiol. 2012; 114(2):457-69. DOI: 10.1111/jam.12061. View

13.

Soo P, Tseng C, Ling S, Liou M, Liu C, Chao H . Rapid and sensitive detection of Acinetobacter baumannii using loop-mediated isothermal amplification. J Microbiol Methods. 2012; 92(2):197-200. DOI: 10.1016/j.mimet.2012.11.020. View

14.

Wang Q, Zhou Y, Li S, Zhuo C, Xu S, Huang L . Real-time fluorescence loop mediated isothermal amplification for the detection of Acinetobacter baumannii. PLoS One. 2013; 8(7):e66406. PMC: 3699609. DOI: 10.1371/journal.pone.0066406. View

15.

Antunes L, Visca P, Towner K . Acinetobacter baumannii: evolution of a global pathogen. Pathog Dis. 2013; 71(3):292-301. DOI: 10.1111/2049-632X.12125. View

16.

Zhou H, Yuan Z, Du Y . Prior use of four invasive procedures increases the risk of Acinetobacter baumannii nosocomial bacteremia among patients in intensive care units: a systematic review and meta-analysis. Int J Infect Dis. 2014; 22:25-30. DOI: 10.1016/j.ijid.2014.01.018. View

17.

De Gregorio E, Roscetto E, Iula V, Martinucci M, Zarrilli R, Di Nocera P . Development of a real-time PCR assay for the rapid detection of Acinetobacter baumannii from whole blood samples. New Microbiol. 2015; 38(2):251-7. View

18.

Wang Y, Wang Y, Ma A, Li D, Luo L, Liu D . Rapid and Sensitive Isothermal Detection of Nucleic-acid Sequence by Multiple Cross Displacement Amplification. Sci Rep. 2015; 5:11902. PMC: 4648395. DOI: 10.1038/srep11902. View

19.

Wang Y, Yan W, Wang Y, Xu J, Ye C . Rapid, sensitive and reliable detection of Klebsiella pneumoniae by label-free multiple cross displacement amplification coupled with nanoparticles-based biosensor. J Microbiol Methods. 2018; 149:80-88. DOI: 10.1016/j.mimet.2018.05.003. View

20.

Wang Y, Yan W, Fu S, Hu S, Wang Y, Xu J . Multiple Cross Displacement Amplification Coupled With Nanoparticles-Based Lateral Flow Biosensor for Detection of and Identification of Methicillin-Resistant . Front Microbiol. 2018; 9:907. PMC: 5954800. DOI: 10.3389/fmicb.2018.00907. View