Organoid as a culture system for viral vaccine strains

Organoid is an in vitro multicellular form mimicking in vivo organ. Its similarity to human organ including cellular organization, molecular expression patterns, as well as genetic signatures enables to study the characteristics of infectious agents and host-pathogen interaction. For the features of organoid, this system also can be potentially used to cultivate currently uncultivable viruses of vaccine candidates. This paper will briefly describe problems in the current culture system for virus production and the possibility of organoid as culture system for viral vaccine and their current limitations that should be solved to meet the goal.

Current status and prospects of laboratory diagnosis of clinical virus infection / 中华检验医学杂志

The severe infectious diseases caused by virus are occurring with increasing frequency, which poses a rising global threat to human health and life.There are many kinds of viruses that cause a wide variety of viral diseases.The same kind of virus is able to cause different diseases, meanwhile a disease can be caused by different viruses.All these conditions make the relationship between viral pathogens and infection diseases complicated.At present, no effective laboratory detection methods for most of virus infectious diseases are developed.But the rapid development of molecular diagnostic technologies makes it possible for clinical laboratory to detect viral infection diseases rapidly and simultaneously.This review summarized the present and development perspectives of laboratory tests for the diagnosis of clinical virus infections, attempting to give some advices for laboratory diagnosis procedure of viral infections.

Characteristics of astrovirus infection and typing characteristics analysis among children hospitalized with diarrhea in Tianjin / 中华检验医学杂志

Objective To study the pathogenic prevalence and genotypes of astrovirus among children under 5 years old hospitalized with diarrhea in Tianjin. Methods A total 837 stool specimens were collected from children with diarrhea hospitalized in Tianjin children's hospital from May 2008 to April 2009. Astrovirus antigens were detected using ELISA and the postive specimens were inoculated in CaCo-2cells. After the CPE caused by virus were observed, the total RNA of virus was extracted, then the genomc fragments of the strains were amplified by using RT-PCR and confirmed by sequencing of the RT-PCR products. Detection of rotavirus was employed by Colloidal Gold Device. Results Astrovirus antigen was found positive in 3.0% of the patients. The coinfection rate of astrovirus and rotavirus was 0. 7% (6/837).Ninety-six persent of children with astrovirus diarrhea were younger than 2 years of age, Forty-eight persent of children with astrovirus diarrhea were younger than 6 months. The astrovirus infections occurred mainly between August 2008 and April 2009. Of the 21 astrovirus positive specimens, 11 cases were successfully identified by RT-PCR and they were all serotype 1. Conclusion Astrovirus is a major cause of nonbacterical diarrhea between 2008 and 2009 in Tianjin, and the predominant serotype is type 1.

Detection and genotyping of rotavirus among children under 5 years old hospitalized with diarrhea in Tianjin / 中华检验医学杂志

Objective To study the prevalence and genotypes of rotavirus (RV) among children,< 5 years old hospitalized with viral diarrhea in Tianjin. Methods Stool specimens were collected from hospitalized diarrhea children in Tianjin children's hospital between May 2008 and April 2009. Detection of rotavirus was employed by Colloidal Gold Device. The detected positives were inoculated to MA-104 cells. The total RNA of virus was extracted after CPE which was caused by rotavirus were observed, The VP7 serotypes were determined by using RT-PCR to amplify the VP7 gene and sequencing the RT-PCR products.The clinical data for each patient were also collected. Results Among 837 specimens, the RV antigen positive rate was 26. 3% (220/837). Among all the children with rotavirus diarrhea, 90. 5% (199/220)were < 2 years old. The prevalence of rotavirus diarrhea in children peaked during Oct. 2008 through Apr.2009. Of the 208 rotavirus positive specimens, 95 were successfully identified by RT-PCR Thirty-five positive strains of RV were sequenced, and the sequencing results showed that 32 positive strains were belonged to rotavirus G1 type, 2 positive strains were belonged to rotavirus G3 type and 1 positive strain were belonged to rotavirus C9 type. Conclusion RV was the dominant etiological agent for infantile diarrhea infection in Tianjin, and the predominant serotype was G1.

Infection kinetics of human adenovirus serotype 41 in HEK 293 cells

The purpose of this work was to acquire an overview of the infectious cycle of HAdV-41 in permissive HEK 293 cells and compare it to that observed with the prototype of the genus, Human adenovirus C HAdV-2. HEK 293 cells were infected with each virus separately and were harvested every 12 h for seven days. Infection kinetics were analysed using confocal and electronic microscopy. The results show that, when properly cultivated, HAdV-41 was not fastidious. It had a longer multiplication cycle, which resulted in the release of complete viral particles and viral stocks reached high titres. After 60 h of infection, the export of viral proteins from the infected cell to the extracellular milieu was observed, with a pattern similar to that previously described for HAdV-2 penton-base trafficking after 30 h of infection. HAdV-41 had a non-lytic cycle and the infection spread from the first infected cell to its neighbours. The release process of the viral particles is unknown. The results observed for HAdV-41 infection in HEK 293 cells show how different this virus is from the prototype HAdV-2 and provides information for the development of this vector for use in gene therapy.

Application of the rapid influenza testing during 2007-2008 flu season in Beijing city / 中华检验医学杂志

Objective To analyze the application value of the rapid testing for influenza during 2007-2008 flu season at fever clinic in Beijing Chaoyang hospital Methods 500 patients with diagnosis of influenza-like illness were prospectively enrolled. Pharyngeal swabs were collected for influenza viral culture and rapid testing for influenza. Demographic characteristics, age, symptoms, lab tests, symptom recovery time and medical expense were also collected. The sensitivity, specificity, positive predictive value and negative predictive value for rapid testing were analyzed. Results A total of 500 patients were enrolled between Dec 2007 and March 2008. Among them 498 cases were used for analysis. Influenza B was most common by virus culture methed(n=208,41.8%) ,followed by influenza A (n=51,10.2%). The average age was 35, and the ratio of male to female was 1.47:1. Compared with the group of positive culture, patients with influenza were more likely to get cough, sore throat, and nasal congestion (t=13.728, 4.014and 4.720,P<0.001 or 0.05, respectively). A total of 260 cases were subjected to rapid testing, Among them 18 cases were influenza A positive and 132 cases were influenza B positive. The rapid testing had a sensitivity of 77.1 % and a specificity of 70.1%. The positive predictive value was 78.6% and the negative predictive value was 68.2%. The rapid testing had enhanced the proportion of anti-viral treatment from 0 to 26% and reduced the proportion of antibiotic use from 63.4% to 20. 7%. Conclusions Influenza B is the most predominant pathogen during 2007-2008 flu season among patients with influenza-like illness in Beijing. The rapid testing with high sensitivity and specificity provides guidance on clinical practice.

Diagnóstico de conjuntivite adenoviral pelo RPS Adenodetector® / Adenovirus conjunctivitis diagnosis using RPS Adenodetector®

OBJETIVO: Avaliar a utilização do RPS Adenodetector®, como método diagnóstico de pacientes com quadro clínico de conjuntivite adenoviral. MÉTODOS: Análise de série de casos consecutivos de pacientes com diagnóstico clínico de ceratoconjuntivite adenoviral submetidos comparativamente ao teste RPS Adenodetector® e a raspado conjuntival para cultura de vírus. RESULTADOS: Dos 11 pacientes avaliados, 10 pacientes apresentavam acometimento unilateral. Em relação ao tempo de início dos sintomas no momento da colheita, 5 (45,5 por cento) pacientes apresentavam dois dias de história, 5 (45,5 por cento) apresentavam três dias e 1 (9,1 por cento) apresentava 7 dias. A cultura para adenovírus foi positiva em 8 pacientes (73 por cento) e o RPS Adenodetector® foi positivo em 9 pacientes (82 por cento). Oito pacientes apresentaram o teste rápido e cultura positiva. Um paciente apresentou teste RPS Adenodetector® positivo com cultura negativa. Os dois pacientes com teste RPS Adenodetector® negativo apresentaram cultura negativa. O RPS Adenodetector® mostrou sensibilidade de 100 por cento e especificidade de 67 por cento adotando-se a cultura de vírus como exame padrão-ouro para o diagnóstico de conjuntivite adenoviral. CONCLUSÃO: O RPS Adenodetector® foi útil para o diagnóstico de conjuntivite adenoviral e pode auxiliar na orientação do paciente quanto ao contágio e disseminação da doença.

PURPOSE: To evaluate the RPS Adenodetector®, a rapid immunochromatographic test, in the diagnosis of patients with clinical overt adenoviral conjunctivitis. METHODS: Consecutive case series. Patients underwent conjunctiva scraping for RPS Adenodetector® test and culture to identify adenovirus. RESULTS: A total of 11 patients were studied, and 10 had unilateral disease. Five (45.5 percent) had symptoms for 2 days, 5 for three days, and 1 for 7 days. Adenovirus culture was positive in 8 patients (73 percent) and RPS Adenodetector® was positive in 9 (82 percent) patients. Eight patients had adenovirus identification by both methods. In one patient the RPS Adenodetector® was positive in contrast to a negative culture. The two patients revealing negative RPS Adenodetector® results also had negative cultures. The sensitivity was 100 percent and the specificity was 67 percent. CONCLUSION: The RPS Adenodetector® is a useful tool in the rapid diagnosis of adenovirus conjunctivitis and may contribute to the spread control of this highly contagious disease.

A clinical perspective research of liver function damage caused by none hepatitis virus / 中国基层医药

Objective To explore the characteristic of disease cause,clinical diagnosis and main tactics of therapy in the temporary liver function damage that is not caused by common hepatitis virus.Methods 573 cases of liver function damage of pediatrics ward in the past 2 years were analyzed and studied about the value of the liver protecting therapy.Results In all 573 cases surfered from liver function damage,418 cases were caused by respiratory infection,accounted for 10.8% of the total patients of the same desease in the same period,101 cases of acute infection of the upper respiratory tract,accounted for 15.6%;54 cases of newborn baby's disease,accounted for 3%.The slight degree damage occupied 492 cases,whether or not taking protecting the liver cures,it hadn't the notable difference on the statistics.The middle to heavy degree damage occupied 135 cases,whether taking protecting the liver cures or not,it had the notable nature difference on the statistics.Conclusion Many common diseases of pediatrics can cause temporary liver function damage.Clinician should take attention to this.It is not necessary to carry out liver protecting therapy in the slight damage cases;And the liver protecting therapy can contribute to the recovering of liver function of the middle to heavy degree damage.

Analysis of viral etiology of sporadic viral meningitis / 中华传染病杂志

0.05).) Conclusions Coxsackie virus is the most common pathogen in patients with viral meningitis in Chaoshan district, and the organization of disease prevention and scientific research and clinical medical should attach importance to it.

Caracterización de los virus de influenza por el método de la inmunoperoxidasa utilizando anticuerpos monoclonales: Montaje y validación

Se aplicó y validó por primera vez en Cuba el método de inmunoperoxidasa para la clasificación rápida en tipo y subtipo de los virus de influenza. El método está basado en un cultivo rápido en células MDCK-L y el empleo de anticuerpos monoclonales para la clasificación en tipo y subtipo. Se utiliza un pool de anticuerpos contra influenza A y otro contra la influenza B y los anticuerpos monoclonales HA1 - 71 y HA2 - 76 para el subtipaje en H1 y H3. La validación se realizó aplicándolo a 21 cepas de referencia internacional y 23 cepas de virus de influenza humana, aisladas y clasificadas previamente por inhibición de la hemaglutinación. Todas las cepas reaccionaron frente a los anticuerpos monoclonales en correspondencia con su tipo y subtipo de hemaglutinina. Fueron caracterizadas 6 cepas de referencia y 9 aislamientos dentro del subtipo H1N1; 9 cepas de referencia y 10 aislamientos en el subtipo H3N2 y 6 cepas de referencias y 4 aislamientos dentro del tipo B.No hubo reacciones inespecíficas ni cruzadas en los controles establecidos. La sensibilidad, especificidad y coincidencia fue del 100 %. La técnica resultó rápida y conveniente para la caracterización en tipo y subtipo de las cepas de virus de Influenza aisladas. Puede sustituir al método clásico de inhibición de la hemaglutinación cuando se requiere la caracterización rápida de brotes o epidemias de infecciones respiratorias agudas, lo que es de extrema importancia por ser éstas causantes de una alta morbilidad, fundamentalmente en grupos de riesgo, y por su repercusión económica.

The immunoperoxidase method for the rapid classification of influenza viruses in type and subtype was applied and validated for the first time in Cuba. The method is based on a rapid culture in MDCK-L cells and on the use of monoclonal antibodies for the classification in type and subtype. A pool of antibodies against influenza A and another against influenza B and HA1-71 and HA2-76 monoclonal antibodies are used for the subtyping in H1 and H3. The validation was carried out by applying this method to 21 international reference strains and to 23 human influenza virus strains that were isolated and previously classified by hemagglutination inhibition. All the strains reacted to the monoclonal antibodies according to their hemagglutinin type and subtype. 6 reference strains and 9 isolations were characterized within the H1N1 subtype: 9 reference strains and 10 isolations in the H3N2 subtype; and 6 reference strains and 4 isolations in type B. There were neither unspecific nor crossed reactions among the controls established. There was 100 % of sensitivity, specificity and coincidence. The technique used proved to be fast and convenient for the characterization in type and subtype of the isolated influenza virus strains. It may substitute the classic hemagglutination inhibition method when it is required the rapid characterization of outbreaks or epidemics of acute respiratory infections, which is very important due to the high morbidity they cause mainly in risk groups and to their economic repercussion.