Enterovirus 71 (EV71) infection is more likely to induce severe complications and mortality than other enteroviruses. Methods for detection of IgM antibody against EV71 had been established for years, however, the performance of the methods in the very early diagnosis of EV71 infection had not been fully evaluated, which is especially meaningful because of the short incubation period of EV71 infection. In this report, the performance of an IgM anti-EV71 assay was evaluated using acute sera collected from 165 EV71 infected patients, 165 patients infected with other enteroviruses, and more than 2,000 sera from healthy children or children with other infected diseases. The results showed a 90% sensitivity in 20 patients who were in their first illness day, and similar sensitivity remained till 4 days after onset. After then the sensitivity increased to 95% to 100% for more than one month. The specificity of the assay in non-HFMD children is 99.1% (95% CI: 98.6–99.4), similar as the 99.9% specificity in healthy adults. The cross-reaction rate in patients infected with other non-EV71 enteroviruses was 11.4%. In conclusion, the data here presented show that the detection of IgM anti-EV71 by ELISA affords a reliable, convenient, and prompt diagnosis of EV71 infection.
In utero exposure to organochlorine pesticides (OCPs) is thought to be potentially harmful to fetal development. We aimed to investigate the associations of maternal and cord serum OCPs levels with infant birth weight in China. In this study, we measured serum levels of 18 OCPs in 81 mother-infant pairs, including DDT, hexachlorocyclohexanes (BHC), hexachlorobenzene (HCB), heptachlors, chlordanes, endosulfan-I, and mirex using a high-resolution-gas-chromatography with high-resolution-mass-spectrometry method. We found that p,p’-DDE and β-BHC had the highest detection rate in both maternal and cord blood serum (97.2% and 96.7%, respectively), followed by HCB (93.0%, 51.7%), p,p’-DDT (88.7%, 36.7%), and p,p’-DDD (83.1%, 60.0%). Among all OCPs, the concentration of p,p’-DDE was the highest (mothers geometric mean (GM): 203.54 ng/g, newborns GM: 116.14 ng/g), followed by HCB (70.62 ng/g, 65.16 ng/g), and β-BHC (67.67 ng/g, 33.39 ng/g). Multiple linear regression analyses showed that each 1 ng/g increment of cord serum p,p’-DDE, total DDT, and β-BHC was associated with a 0.10g, 0.10g, and 0.92g decrease in infant birth weight, respectively, and as the cord serum concentrations of p,p'-DDT, p,p’-DDD, HCB and mirex increased, the infant birth weight was also decreased, although the associations were not statistically significant due to the relatively small sample size. These results suggest that p,p’-DDE, β-BHC, and HCB were the predominant OCPs in the serum of Chinese pregnant women and cord blood of their newborns. Prenatal exposure to DDT, β-BHC, HCB, and mirex were associated with a decrease in birth weight, but these results need validation in larger sample-sized studies.
Armored RNA has been increasingly used as both an external and internal positive control in nucleic acid-based assays for RNA virus. In order to facilitate armored RNA purification, a His 6 tag was introduced into the loop region of the MS2 coat protein, which allows the exposure of multiple His tags on the surface during armored RNA assembly. The His-tagged armored RNA particles were purified to homogeneity and verified to be free of DNA contamination in a single run of affinity chromatography. A fragment of severe acute respiratory syndrome coronavirus (SARS-CoV) genome targeted for SARS-CoV detection was chosen for an external positive control preparation. A plant-specific gene sequence was chosen for a universal noncompetitive internal positive control preparation. Both controls were purified by Co 2؉ affinity chromatography and were included in a real-time reverse transcription-PCR assay for SARS-CoV. The noncompetitive internal positive control can be added to clinical samples before RNA extraction and enables the identification of potential inhibitive effects without interfering with target amplification. The external control could be used for the quantification of viral loads in clinical samples.External positive controls (EPC) and internal positive controls (IPC) are important components in nucleic acid-based testing. An EPC is used to validate or calibrate the established assays, while an IPC is employed to reveal potential inhibition (5,13,15,16,20,25). Ideally, both controls should closely resemble the target of interest so as to track the entire process of the assay, including pathogen lysis, nucleic acid extraction, RNA reverse transcription (RT), amplification, and detection. These controls should also be stable for long-term storage and safe for routine use. Currently used positive controls, such as pure plasmids or naked RNA, cannot fulfill these requirements. In RT-PCR particularly, naked RNA fragments are too labile, whereas plasmid DNA, although more stable than RNA fragments, is not representative of an authentic template in the RT procedure (6).Engineered phages are good alternatives for control preparations and have been successfully used to prepare nucleaseresistant RNA positive controls, armored RNA (AR), for 17,18,24), as well as DNA positive controls (lambda phage DNA) for PCR assays (22). The advantages of phage controls include safety, stability, and physical characteristics that mimic the natural virus. Due to the extreme lability of RNA, AR controls have been widely used (2,3,8,9,14). Unfortunately, only a limited number of AR controls are commercially available so far. Most of the commercial ARs are of little assistance to researchers, since they often use target sequences of their own, which requires customer preparation of special AR. The lack of safe and stable RNA controls could become a serious problem in cases of newly emerging pathogens such as severe acute respiratory syndrome coronavirus (SARS-CoV), where cross-infection could happen during sample exchange between laborat...
Incidence of hepatocellular carcinoma (HCC) ranked the fifth in male and ninth in the female counterparts, and 50% of incidence HCC cases were occurred in China with high hepatitis B virus (HBV) prevalence. HCC has seriously compromised the health status of general population in China. A case–control study of 314 HCC cases and 346 controls was conducted in Xiamen, which is an epidemic area in China for both hepatitis B infection and HCC. Face-to-face interview was conducted to gather information on demographic characteristics as well as exposure of environmental factors. Commercial enzyme-linked immunosorbent assay kits were used to determine the status of serological markers of HBV infection. Odds ratios and 95% confidence intervals were estimated by using unconditional logistic regression. Multivariate unconditional logistic regression analysis was applied to evaluate the potential interactions of variables or confounders.As expected, HBV and alcohol intake still are the major risk factors of HCC. Liver disease history and passive smoking are also associated with elevated HCC risk. Indoor air pollution and pesticide exposure have newly identified as risk factors of HCC. Fruit and tea intake can significantly lower the HCC risk.The application of HBV vaccine and reduction on alcohol intake should be further promoted in high-risk population. Fruit and tea can be served as chemoprevention in daily life due to their high accessibility.
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