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The recent pneumonia outbreak caused by a novel coronavirus (SARS-CoV-2) is posing a great threat to global public health. Therefore, rapid and accurate identification of pathogenic viruses plays a vital role in selecting appropriate treatments, saving people's lives and preventing epidemics. It is important to establish a quick standard diagnostic test for the detection of the infectious disease (COVID-19) to prevent subsequent secondary spread. Polymerase chain reaction (PCR) is regarded as a gold standard test for the molecular diagnosis of viral and bacterial infections with high sensitivity and specificity. Isothermal nucleic acid amplification is considered to be a highly promising candidate method due to its fundamental advantage in quick procedure time at constant temperature without thermocycler opera-tion. A variety of improved or new approaches also have been developed. This review summarizes the currently available detection methods for coronavirus nucleic acid. It is anticipated that this will assist researchers and clinicians in developing better techniques for timely and effective detection of coro-navirus infection.
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Objective:To analyze the viral nucleic acid and cytokines in 12 children with 2019-nCoV infection.Methods:Clinical and laboratory data of the children diagnosed with 2019-nCoV infection in Guangzhou Women and Children′s Medical Center from January to April 2020 were retrospectively analyzed. Throat and anal swabs were collected on alternate days for the detection of 2019-nCoV nucleic acid by fluorescence quantitative PCR. Flow cytometry was used to detect serum cytokines including IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12p70, IL-17A, IL-17F, IL-22, TNF-α and TNF-β during the early (both throat and anal swab tests were positive), the intermediate (throat swab test was negative, while anal swab test remained positive), and the convalescence (both throat and anal swab tests were negative) stages of infection.Results:A total of 12 children were enrolled in this study. The male-to-female ratio was 5∶1. The average age was (7.0±4.3) years. There were two asymptomatic, five mild and five common cases. No severe or critical cases were involved. Initially, throat and anal swab nucleic acid tests were simultaneously positive in nine children newly diagnosed in our hospital and the median time of viral shedding in throat swab was longer than that in throat swab [32 (4.5, 45.0) d vs 3 (2, 9) d, Z=11.0, P=0.010]. The median difference of viral shedding time between anal swab and pharyngeal swab was 25.5 (1.5, 42.8) d. The overall levels of serum cytokines IL-17A, IL -4 and IL-5 in different stages of the disease (early, intermediate and convalescence stage) were statistically different ( Z or F, P values were 8.33, 0.016; 5.36, 0.010 and 6.56, 0.004, respectively), and a significant increase was observed in the intermediate stage of infection. IL-17F, IL-2 and IL-22 were all increased during the infection, but there was no significant statistical difference among the three stages ( P>0.05). Conclusions:It was noted that intestinal viral shedding needed a longer time. Although the infectivity has not been determined, higher requirements have been put forward for disease prevention and control. Cytokines secreted by Th2 and Th17 cells were involved in the immune response in children with non-severe 2019-nCoV infection. Monitoring viral shedding and cytokine changes in pediatric patients would be conducive to disease assessment.
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Objective@#To investigate the clinical manifestations, diagnosis, and treatment of H1N1 influenza A-associated encephalopathy (IAE) in children.@*Methods@#The clinical manifestations, laboratory tests, cranial magnetic resonance imaging (MRI), electroencephalography (EEG) examinations and treatments of seven children with H1N1 IAE hospitalized in Guangzhou Women and Children′s Medical Center from December 2018 to January 2019 were retrospectively analyzed.@*Results@#Five of the seven children with H1N1 IAE were female. The age at admission was 4 years and 5 months (range 7 months-9 years). Neurological symptoms occurred simultaneously or early (0-3 days) after the flu-like symptom appeared. The main clinical manifestations of neurological symptoms were seizures (repeated seizures in five cases and status convulsion in two cases, including one case of unexpected fever and repeated seizures in a nine-year old girl) accompanied with altered consciousness (drowsiness in five cases and coma in two cases). Cranial MRI in three cases displayed multifocal lesions, mainly in the bilateral thalamus, brainstem and cerebellar hemisphere. MRI also showed reversible splenial lesion in the corpus callusumin in three cases. EEG tracings were characterized by diffuse slow wave activity in four cases, and status epilepticus was monitored in one case. All the 7 cases were treated with oral oseltamivir. Three cases were treated with pulsed methylprednisolone and intravenous immunoglobulin. One case was treated with intravenous immunoglobulin alone and all the patients received oral oseltamivir. All the patients survived, with three patients had minor neurological sequelae at discharge.@*Conclusions@#The main clinical manifestations of H1N1 IAE are seizures and altered consciousness. Cranial MRI combined with EEG is helpful for early diagnosis. Intravenous immunoglobulin and (or) methylprednisolone should be considered for severe cases.
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MicroRNAs (miRNAs) are a family of endogenous, small (approximately 22 nucleotides in length), noncoding, functional RNAs. With the development of molecular biology, the research of miRNA bio-logical function has attracted significant interest, as abnormal miRNA expression is identified to contribute to serious human diseases such as cancers. Traditional methods for miRNA detection do not meet current demands. In particular, nanomaterial-based methods, nucleic acid amplification-based methods such as rolling circle amplification (RCA), loop-mediated isothermal amplification (LAMP), strand-displacement amplification (SDA) and some enzyme-free amplifications have been employed widely for the highly sensitive detection of miRNA. MiRNA functional research and clinical diagnostics have been accelerated by these new techniques. Herein, we summarize and discuss the recent progress in the development of miRNA detection methods and new applications. This review will provide guidelines for the development of follow-up miRNA detection methods with high sensitivity and spec-ificity, and applicability to disease diagnosis and therapy.
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Objective To explore the clinical characteristics of children with dengue fever (DF) hospitalized in Guangzhou in 2014 , and to raise clinician′s level of understanding of dengue fever in children.Methods Clinical data of 78 children hospitalized with DF in the Department of Infectious Diseases ,Guangzhou Women and Children′s Medical Center were retrospectively analyzed .Chi-square was used for discontinuous variables ,and t test was used for continuous variabbles .Results The 78 cases aged 27 days to 14 years old ,with median of 5 years old .Fifty cases (64 .1%) were male ,and 28 cases (35 .9%) were female.Epidemic areas had gathering trend ,mainly in central urban area .Major clinical manifestations were fever (100 .00%) , rash (82 .05%) , myalgia／fatigue (28 .21%) , but without diarrhea ,jaundice ,hematemesis or hematochezia .Laboratory tests suggested leukopenia (80 .77%) and thrombocytopenia (82 .05%) ,abnormal blood coagulation function with prolonged APTT (57 .69%) ,and abnormal liver function (47 .44%).Etiology examinations showed 66 cases of children had dengue virus nucleic acid detected 1－10 days after onset ,with the positive rate of 89 .19%(66／74).A total of 48 cases had IgM positive ,with the positive rate of 81 .36%(48／59).IgM began to appear as early as the first day of disease onset ,and the average period was (5 .5 ± 0 .8) days .Dengue virus type 1 was the main type . Conclusions In 2014 , dengue virus type 1 is the main strain causing dengue fever in children in Guangzhou .Fever ,rash ,leukopenia ,thrombocytopenia ,clotting disorders and liver function damage are the main clinical features .No serious or fatal cases are reported ,and the prognosis is good.
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Objective To investigate the efficacy and safety of domestic Oseltamivir granules in the treatment of influenza A in children.Methods The research subjects were the hospitalized patients with influenza A coming from the Department of Infectious Disease in Guangzhou Women and Children's Medical Center.from January and March,2014 They were divided into 2 groups randomly.Patients in the treatment group received domestic Oseltamivir granules,while the patients in the control group received foreign Oseltamivir.Chi square test was performed to compare baseline characteristics and the difference in side effects.Paired t test was used to compare baseline characteristics and the difference in side effects.Results There were 44 cases in the study group,27 cases were male(61.36％),17 cases were female (38.64％),and the average age was (2.40 ± 1.97) years old.There were 40 cases in the control group,28 cases were male (70.00％),12 cases were female (30.00％),and the average age was (2.99-± 3.19) years old.The maximum age was 6 years old.The age and gender were not significant (P ＞ 0.05) between 2 groups.The fever remission time was (14.67-± 9.37) h and (11.80-± 9.98) h in the treatment group and the control group,and there was no significant difference between 2 groups (P ＞ 0.05);the flu symptoms completely remission time was (62.50 ± 25.03) h and (65.40 ± 36.74) h in the treatment group and the control group,and there was no significant difference between 2 groups (P ＞ 0.05);the adverse reaction was not observed in the treatment group,but 1 case in the control group had cough,and without special treatment cough alleviated gradually;the close contacts in 2 groups did not develop influenza A.Conclusions The domestic Oseltamivir granules are as effective and safe as the foreign Oseltamivir.And the domestic Oseltamivir granules are the granular type,and therefore more suitable for children