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1.
Sci Rep ; 13(1): 18781, 2023 10 31.
Article in English | MEDLINE | ID: mdl-37907677

ABSTRACT

Uropathogenic Escherichia coli (UPEC) causes up to 90% of urinary tract infections (UTI) which is more prevalent among females than males. In urine, patients with symptomatic UTI usually have a high concentration of bacterial infection, ≥ 105 colony-forming units (CFU) per mL, in which the culture method is regularly the gold standard diagnosis. In this study, a simple and inexpensive distance-based paper device (dPAD) combined with the fluorescent closed tube LAMP assay was validated for simultaneously screening and semi-quantifying the infection level of E. coli in 440 urine samples of patients with UTI. The dPAD could measure the LAMP amplicons and semi-quantify the levels of E. coli infection in heavy (≥ 104 CFU/mL), light (≤ 103 CFU/mL) and no infection. The sensitivity and specificity had reliable performances, achieving as high as 100 and 92.7%, respectively. The one step LAMP assay could be performed within 3 h, which was 7.5 times faster than the culture method. To empower early UTI diagnosis and fast treatment, this inexpensive dPAD tool combined with the fluorescent closed tube LAMP assay is simple, reliably fast and practically portable for point-of-care settings, particularly in resource-limited areas, which can be set up in all levels of healthcare facilities.


Subject(s)
Escherichia coli Infections , Urinary Tract Infections , Female , Humans , Escherichia coli/genetics , Nucleic Acid Amplification Techniques/methods , Molecular Diagnostic Techniques , Escherichia coli Infections/diagnosis , Escherichia coli Infections/microbiology , Urinary Tract Infections/diagnosis , Urinary Tract Infections/microbiology
2.
Analyst ; 145(24): 8077-8086, 2021 Jan 07.
Article in English | MEDLINE | ID: mdl-33078771

ABSTRACT

The highly sensitive and selective determination of Escherichia coli (E. coli) in urine was achieved using a SYBR™ safe loop-mediated isothermal amplification (LAMP) method with a distance-based paper device. New primers set specific to multi-copy the 16s rRNA gene of E. coli were designed and used in this study. The detection sensitivity of these primers was higher than in related work and they could be incorporated with a low-cost paper-based device to quantify E. coli in urine at a concentration lower than 101 CFU mL-1. Regarding standard artificial urine, a linear range of a 10-fold dilution of E. coli concentration (105-100 CFU mL-1) with an R-square value (R2) = 0.9823 was observed directly using a fluorescent migratory distance of the 4 µL reaction mixture in the detection zone under blue light without the need for postreaction staining process. Based on the device, E. coli infection could be significantly categorized into 3 groups; none, light, and heavy levels, which is beneficial for UTI diagnosis. Hence, this paper-based device is suitable for use with the SYBR™ Safe-LAMP assay to semi-quantify E. coli, especially in resource-limited settings due to advantages of low cost, simple fabrication and operation, and no requirement for sophisticated instruments, as well as its disposability and portability.


Subject(s)
Escherichia coli , Nucleic Acid Amplification Techniques , Escherichia coli/genetics , Molecular Diagnostic Techniques , RNA, Ribosomal, 16S/genetics , Sensitivity and Specificity
3.
J Med Microbiol ; 63(Pt 5): 753-759, 2014 May.
Article in English | MEDLINE | ID: mdl-24591706

ABSTRACT

Oral candidiasis is a common opportunistic infection among human immunodeficiency virus (HIV)-infected individuals, with growing concerns about the emergence of non-albicans species with resistance to antifungal agents. This cross-sectional study determined the prevalence of oral Candida species in Thai HIV-infected adults and factors affecting their colonization. Candida species were identified from oral rinse samples of 60 HIV-infected participants of the MTCT-Plus initiative and 49 healthy controls by culture-based and molecular assays. The prevalence of oral Candida carriage was similar in HIV-infected patients (56.6 %) and in controls (55.1 %, P = 0.87). Candida albicans was the most predominant species in both groups (94.1 % of Candida carriers in HIV, 88.9 % in control). Interestingly, Candida dubliniensis was the second most common species in controls (29.6 %) and the third in HIV-infected patients (11.8 %, P = 0.08). Multivariate analysis showed that, amongst HIV-infected individuals, CD4 count <200 cells mm(-3) was associated with increased prevalence of oral carriage of both C. albicans (P = 0.03) and non-albicans species (P = 0.03). Moreover, patients with tuberculosis infection had a higher prevalence of the non-albicans species than those without (P = 0.03). Intriguingly, contraceptive use was also associated positively with non-albicans and multi-species carriage (P = 0.04 for both). However, use of antiretroviral drugs protected the patients from Candida carriage (P = 0.03), especially from C. albicans (P = 0.02). In conclusion, while HIV-infected individuals had a similar prevalence of oral Candida carriage to that of the control group, host immune status, tuberculosis infection, and contraceptive use may influence oral colonization of Candida, especially of the non-albicans species.


Subject(s)
Candida/isolation & purification , Candidiasis, Oral/epidemiology , Candidiasis, Oral/microbiology , Carrier State/epidemiology , Carrier State/microbiology , HIV Infections/complications , HIV Infections/immunology , Adult , Anti-Retroviral Agents/therapeutic use , CD4 Lymphocyte Count , Candida/classification , Cross-Sectional Studies , Female , HIV Infections/drug therapy , Humans , Male , Middle Aged , Prevalence , Thailand/epidemiology , Tuberculosis/complications , Young Adult
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