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

Abstract

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/μL, respectively, and C. trachomatis was 41.3 copies/μL; on LAMP-microfluidic chip they were 260, 154, 3.9 and 7.53 CFU/μL, respectively, and C. trachomatis was 120 copies/μL. 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.

The Potential Clinical Applications of a Microfluidic Lab-on-a-Chip for the Identification and Antibiotic Susceptibility Testing of Enterococcus faecalis-Associated Endodontic Infections: A Systematic Review

“…Following a thorough examination of the complete texts, an additional 16 studies were excluded as they did not align with the predefined selection criteria. Ultimately, this systematic review incorporated 12 studies [21][22][23][24][25][26][27][28][29][30][31][32], as illustrated in Figure 1. The characteristics of the studies encompassed in the analysis are detailed in Table 1.…”

“…The PCR amplification of the ex-The genomic DNA of E. faecalis The characteristics of the studies encompassed in the analysis are detailed in Table 1. The publications incorporated in this review span the timeframe from 2009 [21] to 2022 [31,32].…”

“…Table 1 shows that out of the 12 studies assessed in this systematic review, 7 aimed to identify E. faecalis [21,24,25,27,29,31,32], while the remaining 5 evaluated its susceptibility to different antibiotics [22,23,26,28,30]. In all the studies, it was observed that the newly developed microfluidic chip significantly reduces the detection time when compared to traditional methods.…”

A loop-mediated isothermal amplification-enabled analytical assay for the detection of SARS-CoV-2: A review

“…The development of a lab-on-chip (LOC) system based on the integration of LAMP and microfluidic technology is expected to speed the detection of pathogens and allow the early diagnosis of diseases (Figure 3). Currently, LAMP technology on LOC has been applied to the detection of breast cancer biomarkers, dengue serotypes, carbapenemase-producing organisms, and multiple pathogens in the lower reproductive tract during pregnancy (Moser et al, 2022;Wormald et al, 2022;Wu et al, 2022;Xu et al, 2022). Microfluidic chips are characterized by high throughput and low cost, a method of LAMP technology combined with a microfluidic chip to detect a Li et al 10.3389/fcimb.2022.1068015 variety of pathogens has been established, which can perform detection of multiple pathogens and real-time detection within 40 min without aerosol pollution and cross reaction (Ou et al, 2021).…”

A microfluidic-chip-based system with loop-mediated isothermal amplification for rapid and parallel detection of Trichomonas vaginalis and human papillomavirus

“…25,26 In recent years, the use of microfluidic chips has significantly enhanced the capabilities of LAMP assays by enabling high-throughput and simultaneous detection of multiple targets in a reagent-efficient and user-friendly manner. 27,28 Additionally, recent studies have demonstrated the successful application of LAMP assays in identifying T. vaginalis or HPV. 23,29 Hence, by combining LAMP and microfluidic technology, it becomes possible to achieve parallel and rapid detection of HPV and T. vaginalis, which can be instrumental in managing the risk of cervical cancer.…”