Rapid and simultaneous detection of multiple pathogens in the lower reproductive tract during pregnancy based on loop-mediated isothermal amplification-microfluidic chip

被引:10
作者
Xu, Xiaofang [1 ]
Jia, Yiguo [1 ]
Li, Ruolin [1 ]
Wen, Yuting [2 ]
Liang, Yuchen [3 ]
Lao, Guangjie [1 ]
Liu, Xiaojuan [4 ]
Zhou, Wei [5 ]
Liu, Huawei [6 ]
Xie, Jiang [6 ]
Wang, Xiaoxia [7 ]
Xu, Wenming [2 ]
Sun, Qun [1 ]
机构
[1] Sichuan Univ, Coll Life Sci, Key Lab Bioresources & Ecoenvironm, Minist Educ, Chengdu 610065, Sichuan, Peoples R China
[2] Sichuan Univ, West China Univ Hosp 2, Dept Obstet Gynecol,Minist Educ, Joint Lab Reprod Med,SCU CUHK,Key Lab Obstetr Gyn, Chengdu 610041, Peoples R China
[3] Shanghai Jiao Tong Univ, Sch Life Sci & Biotechnol, Shanghai, Peoples R China
[4] Sichuan Univ, West China Univ Hosp 2, Dept Lab Med, Chengdu, Peoples R China
[5] Sichuan Univ, Key Lab Birth Defects & Related Dis Women & Child, Minist Educ, Chengdu, Peoples R China
[6] Southwest Jiao Tong Univ, Peoples Hosp Chengdu 3, Affiliated Hosp, Chengdu, Peoples R China
[7] Tianjin Chase Sun Pharmaceut Co Ltd, 20 Quanfa Rd, Tianjin 301700, Peoples R China
关键词
Reproductive tract infection; Pathogen; Loop-mediated isothermal amplification; Microfluidic technology; LAMP-microfluidic chip; Rapid and simultaneous detection; GARDNERELLA-VAGINALIS; SEQUENCE; GENE; IDENTIFICATION; INFECTIONS; CHLAMYDIA;
D O I
10.1186/s12866-022-02657-0
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Background Female reproductive tract infection (RTI) is the common source of varied diseases, especially as an important risk factor for pregnancy outcomes, therefore the rapid, accurate and simultaneous detection of multiple pathogens is in urgent need for assisting the diagnosis and treatment of RTI in pregnant women. Streptococcus agalactiae (S. agalactiae), Enterococcus faecalis (E. faecalis), Gardnerella vaginalis (G. vaginalis), Candida albicans (C. albicans) and Chlamydia trachomatis (C. trachomatis) are five main pathogens in lower genital tract with high risk, serious consequences and clinical demands. The combination of loop-mediated isothermal amplification (LAMP) and microfluidic technology was used to develop the LAMP-microfluidic chip for rapid, simple, sensitive and simultaneous detection of the five target pathogens above. Results Standard strains and clinical isolates were used for the establishment of the novel LAMP method in tube and LAMP-microfluidic chip, followed by the chip detection on 103 clinical samples and PCR verification partially. The sensitivities of LAMP of S. agalactiae, E. faecalis, G. vaginalis, and C. albicans in tube were 22.0, 76.0, 13.2, 1.11 CFU/mu L, respectively, and C. trachomatis was 41.3 copies/mu L; on LAMP-microfluidic chip they were 260, 154, 3.9 and 7.53 CFU/mu L, respectively, and C. trachomatis was 120 copies/mu L. The positive coincidence rates of clinical stains in tube and on chip experiments were 100%. Compared with the classic culture method performed in hospitals, the positive coincidence rate of the 103 clinical samples detected by LAMP-microfluidic chip were 100%. For the six inconsistent ones, including four G. vaginalis and two C. albicans positive samples tested by LAMP-microfluidic chip and verified by PCR were negative by culturing method in hospitals, indicating the lack of efficient detection by the classic culturing method. Conclusion Our study suggested that the LAMP-microfluidic chips could simultaneously, efficiently, and accurately detect multiple main pathogens, including S. agalactiae, E. faecalis, G. vaginalis, C. albicans and C. trachomatis, in clinical samples of female RTI to give a great clinical value. Accordingly, this novel method has the potential to provide a valuable reference for female RTI screening and early diagnosis during pregnancy.
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页数:16
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