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The occurrence of Giardia and Cryptosporidium (oo)cysts in drinking source water poses a serious public health risk. Here, we established a method that combines membrane concentration and real-time polymerase chain reaction (PCR) to quantify Giardia and Cryptosporidium in drinking water. The water samples were filtered through a cellulose membrane to collect Giardia and Cryptosporidium, and then nucleic acids were extracted. Specific primers and probes were designed and synthesized according to the gph gene sequence of Giardia and 18S rRNA gene sequence of Cryptosporidium. The concentrations of the two targets were determined using real-time PCR technology. The sensitivity, specificity, and stability of the method were evaluated. Our findings revealed that the detection limits of real-time PCR method for detecting Giardia and Cryptosporidium were 0.926 and 0.65 copy/µL, respectively; the spiked recovery rates were above 60% and 38%, respectively, and relative standard deviations were under 0.95% and 2.26%, respectively. Therefore, this effective procedure based on the membrane concentration method and real-time PCR will be useful for detecting Giardia and Cryptosporidium in drinking water for purpose of continuous environmental monitoring.
Assuntos
Criptosporidiose , Cryptosporidium , Água Potável , Humanos , Cryptosporidium/genética , Giardia/genética , Reação em Cadeia da Polimerase em Tempo RealRESUMO
Objective To analyze serotype distribution, drug resistance, quinolone resistance gene carrying status and genetic relationship of foodborne Salmonella and human Salmonella isolates in Changzhou from 2012 to 2018, to provide scientific basis for the prevention and control of Salmonella. Methods The serum type was identified by serum agglutination and liquid chip. The antibiotic sensitivity was determined by micro broth dilution method. The quinolone antibiotic resistance gene was determined by gene sequencing method. The multilocus sequence typing ( MLST ) typing was performed on quinolone-resistant Salmonella, and the genetic relationship was analyzed by BioNumerics 8.0. Results A total of 10 and 36 serotypes were detected in 46 foodborne Salmonella strains and 152 human Salmonella strains, respectively. The dominant serotypes were Indiana Salmonella and Salmonella typhimurium. Erythromycin resistance rate was the highest in both Salmonella strains, and the proportion of multidrug-resistant bacteria was 93.47 % ( 43 / 46 ) and 80.92 % ( 123 / 152 ), respectively. 38 strains of quinolone-resistant foodborne Salmonella GyrA subunit mainly occurred double mutations Asp87Asn, Ser83Phe, ParC subunit mainly occurred single mutation Ser80Arg, 119 strains of quinolone-resistant human Salmonella qnrS gene detection rate was higher, reached 68.1 % ( 81 / 119 ) ; The dominant ST types of quinolone-resistant Salmonella from two sources were ST17 and ST19, respectively. Conclusions The antibiotic sensitivity of the two Salmonella resistant strains from Changzhou was the same ; Synergistic drug resistance, but both quinolone resistance genemutations and carry inconsistent ; The ST type distribution of quinolone resistant strains isalso inconsistent, and the genetic relationship is far. It is suggested that the probability of Salmonella resistant bacteria infection caused by food transmission in our region is small, and the treatment of the two should be differentiated.
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Objective To probe into the feasibility and applicability of the foodborne food risk monitoring and sentinel hospital foodborne disease surveillance on the Salmonella serotype identification by liquid phase suspension array. Methods The serotyping of Salmonella isolates was performed by traditional glass agglutination test. Meanwhile, the liquid phase suspension array was operated to analyze the antigen O, H and AT for classification identification. Results From 2012 to 2019, a total of 215 strains of Salmonella were collected divided into 38 serotypes, 96% of them could be analyzed by SSA kit. The results of xMAP were in accordance with the traditional agglutination. 8 of the 11 strains which cannot be checked out by glass agglutination seemed to be easy detected by liquid phase suspension array. Conclusion The liquid phase suspension array has advantages of high throughput, high sensitivity and high specificity. It is able to detect the Salmonella serotype rapidly during a short time. Compared with the traditional serum agglutination method, the liquid phase suspension array has obvious advantages in detection time. It can be useful and important in the break out of foodborne disease caused by Salmonella spp.
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Objective To understand the epidemiological characteristics and variation of H3N2 influenza virus hemagglutinin (HA) gene in Changzhou from 2017 to 2018. Methods Throat swab specimens of the influenza-like cases were collected from Changzhou Influenza Monitoring Sentinel Hospital from April 2017 to March 2018. RNA was extracted from the specimens for influenza diagnosing and genotyping using real-time RT-PCR．H3N2 positive samples were isolated, and extracted RNA was used for amplification, sequencing and phylogenetic analysis of HA gene. Results From April 2017 to March 2018, 28 strains of influenza A (H3N2) virus were isolated. After gene sequencing, a phylogenetic tree was constructed. It was found that all of the strains belonged to Group3C.2a, which was similar to the vaccine strain A/Hong Kong/4801/2014. The HA amino acid sequence difference was analyzed and compared between the H3N2 influenza virus strains isolated in Changzhou and the vaccine strain A/Hong Kong/4801/2014. It was found that the epidemic strain isolated in Changzhou was in the HA epitope (A-E) region. Ten amino acid site mutations in the HA epitope (A-E) region and two amino acid site mutations in the stem region of HA antigen were found. Conclusion From April 2017 to March 2018, the influenza virus H3N2 prevalent in Changzhou was distributed on the same evolutionary branch with the vaccine strain A/Hong Kong/4801/2014 (group 3C.2a), rendering the popular trend of one subgroup. However, some amino acid sites of the HA epitope had variations, suggesting that mutations may occur, which may affect the immune effect of the vaccine. Monitoring needs to be strengthened in the future work.