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Adenovirus surveillance on children hospitalized for acute lower respiratory infections in Chile (1988-1996).

Adenoviruses (Ad) play an important role in the etiology of acute lower respiratory tract infections (ALRI) in young children in Chile. Our aim was to correlate the clinical severity of the infections with the Ad strains isolated during surveillance over 8 years. From 1988 through 1996, nasopharyngeal aspirates (NPA) were obtained for viral isolation and immunofluorescence assay (IFA) from children under 2 years of age hospitalized for ALRI; Ad isolates were further studied by restriction enzyme analysis of genomic DNA. Of 3,097 cases enrolled, the Ad isolation rate was 12.6%. The most common admission diagnoses among Ad-positive cases were pneumonia and wheezing bronchitis (69.8%). Duration of Ad shedding was studied in 74 cases by IFA. Children excreting Ad for 4 or more days had a longer hospital stay than those shedding for 1-3 days (mean: 16.8 and 7.2 days, respectively; P <.01). Viral shedding for more than 3 days was associated with more severe outcomes. Genome typing of 221 out of 390 Ad isolates resulted in 87 subgenus C and 134 subgenus B strains, including 123 Ad genome type 7h (55.6%, P <.01). The IFA from the NPA was more sensitive for the detection of subgenus B (51. 5%) than subgenus C infections (24.1%, P <.01). Children shedding Ad 7h had longer hospital stays (P <.01), a higher frequency of rectal temperatures over 39 degrees C (P <.01), and greater need for additional oxygen (P <.02) than subgenus C cases. Four cases requiring mechanical ventilation were associated with Ad 7h infections. The data presented show that, in children hospitalized for ALRI, the genome type 7h was associated with a more severe clinical outcome.

Adenoviridae↗

[The prevalence of Chlamydia trachomatis and Ureaplasma urealyticum cervical infection in infertility women and the observation of therapeutic efficacy].

OBJECTIVE: To study the prevalence of Chlamydia trachomatis (CT) and Ureaplasma urealyticum (Uu) cervical infection in infertility women, and therapeutic effectiveness of tetracycline and qianglimycin. METHOD: CT in cervical swab specimen was detected by cell culture, polymerase chain reaction(PCR) and immunofluorescent assay(IFA), Uu in cervical swab specimen was detected by culture, in a group of 145 infertility women, before and after treatment, and 45 women at productive age who attended obstetric and gynecologic clinic. RESULT: The positive rate of CT by cell culture, PCR, IFA(shell) and IFA(direct smear) was 62.7%, 66.8%, 64.8% and 36.5%, respectively, which was obviously higher than that of the control (P < 0.01). The positive rate of Uu was 33.1%, that of CT and Uu co-infection was 18.6%. Both were obviously higher than those of the control (P < 0.01). CONCLUSION: This study indicates that the prevalence of CT and Uu cervical infection in infertility women is high. CT and Uu cervical infection is closely related to female infertility. The therapeutic effectiveness of tetracycline and qianglimycin is not ideal.

Adult↗

Effect of mouse strain and age on detection of mouse parvovirus 1 by use of serologic testing and polymerase chain reaction analysis.

BACKGROUND AND PURPOSE: Detection of mouse parvovirus 1 (MPV) depends on use of serologic and polymerase chain reaction (PCR) assays. These assays were evaluated for their ability to detect virus-specific antibodies or viral DNA in multiple strains and ages of mice inoculated with MPV. METHODS: Twelve-week-old ICR, BALB/c, C3H, C57BL/6, and DBA/2 mice and four- and eight-week-old ICR mice were inoculated with MPV. Serum was harvested four weeks after inoculation and analyzed by use of recombinant non structural protein 1 (rNS1) enzyme-linked immunosorbent assay (ELISA), minute virus of mice (MVM) ELISA, and MPV indirect fluorescent antibody (IFA), MVM IFA, and MPV hemagglutination inhibition (HAI) assays. Select tissues were harvested and analyzed by use of an MPV-specific PCR assay. RESULTS: The number of mice in each group with detectable MPV-specific antibodies or MPV DNA varied with mouse strain, mouse age when inoculated, and viral dose. Seroconversion in mice inoculated at 12 weeks of age was detected almost exclusively by use of the MPV IFA and MPV HAI assays, whereas seroconversion in almost all mice inoculated at 4 and 8 weeks of age was detected by use of all immunoassays except the MVM ELISA. Viral DNA was detected by use of PCR analysis in all strains and ages of mice except DBA/2 mice. CONCLUSIONS: Mouse strain and age have important roles in seroconversion to nonstructural and structural MPV antigens and persistence of viral DNA in mouse tissues. Therefore, diagnostic serologic testing and PCR analysis should be considered within the context of mouse strain and age at the time of MPV exposure, especially when sentinel mice are used for surveillance.

Animals↗

Human immune response to Puumala virus glycoproteins and nucleocapsid protein expressed in mammalian cells.

Puumala hantavirus (PUUV) glycoproteins G1 and G2 and nucleocapsid protein (N) were expressed in BHK-21 cells by transfection of a plasmid producing a recombinant alphavirus replicon. Coexpression of G1 and G2 from separate constructs seemed to be important for the optimal folding of the glycoproteins, as evaluated by a panel of MAbs detecting conformational epitopes. To evaluate the human antibody response against recombinant G1, G2 and N, several panels of sera were tested by immunofluorescence assay (IFA). Also human sera showed the best reactivity towards G1 and G2 coexpressed from separate transcripts (G1 + G2). Notably, only 2% of the acute sera (total number = 133) contained IgG antibodies against G1 + G2, whereas of old-immunity sera (total number = 100) 87% were G1 + G2 positive. Analysis of a panel of serial patient sera showed that as the immunity matured, IgG antibodies against the recombinant glycoproteins appeared and the titers increased in the course of time, while antibodies against the recombinant N were present already in the acute phase in high titers. The granular fluorescence pattern in PUUV IgG-IFA, associated with the acute phase of immunity, was linked to the presence of antibodies against N, whereas the diffuse fluorescence pattern associated with old-immunity, was linked to the development of antibodies against G1 + G2. The granular fluorescence pattern in PUUV IgG-IFA had a predictive value of 100% for acute PUUV infection. Weak cross-reaction with PUUV glycoproteins was observed in 36% of old-immunity DOBV-specific human sera.

Adolescent↗

[Anaerobic infection and its rapid detection in surgical patients].

OBJECTIVE: To investigate anaerobic infection and its rapid detection in surgical patients. METHODS: 372 specimens were collected from surgical patients for anaerobe detection. To find a rapid detectable method, 110 specimens were collected for comparative studies by using indirect immunofluorescent antibody technique (IFA), enzyme labeled antibody technique (ELAT) and general cultural method (CM). RESULTS: The isolation rate of anaerobes was rather high in pyothorax (88.9%), abdominal abscess (86.7%), appendicitis (83.9%) and cerebral abscess (all five cases were positive). In burn wounds, the rate was about 23.6% especially in deep burn wounds. Among 212 strains detected, the predominant anaerobes were as follows B. melaninogenicus (28.8%), B. fragilis (25%), F. nucleatum (7.5%), C. perfringens (7.5%), and peptostreptococcus (5.7%); most of them (87.8%) were non-sporing anaerobes. Mixed infection with aerobes accounted for 64%. The survey of rapid detection was conducted in B. fragilis, B. melaninogenicus, C. perfringens, F. nucleatum and peptostreptococcus. The detectable rate was higher in IFA (79.1%) and ELAT (80.9%) than in CM (37.3%). CONCLUSION: IFA and ELAT might be rapid, specific, sensitive, and simple for detecting anaerobes.

Abdominal Abscess↗

T-cell responses against tyrosinase 368-376(370D) peptide in HLA*A0201+ melanoma patients: randomized trial comparing incomplete Freund's adjuvant, granulocyte macrophage colony-stimulating factor, and QS-21 as immunological adjuvants.

We conducted a randomized trial in HLA*A0201+ patientswith American Joint Committee on Cancer stage III or IV melanoma immunized with tyrosinase 368-376(370D) peptide and gp100 209-217(210M) peptide to compare the potency of three different adjuvants. Patients received 3 monthly immunizations with 500 microg of each peptide either with incomplete Freund's adjuvant (IFA), QS-21, or granulocyte macrophage colony-stimulating factor (GM-CSF). The primary end point was induction of IFN-gamma release by CD8+ T cells against tyrosinase and gp100 peptides measured by enzyme-linked immunospot assays without in vitro prestimulation measured pretreatment, 2 and 8 weeks after the third vaccination. Four of 9 and 4 of 8 patients immunized using QS-21 and GM-CSF, respectively, developed increased frequencies of CD8+ T cells against tyrosinase 370D peptide compared with 0 of 9 patients immunized using IFA (P = 0.045). T-cell responses against a gp100-related peptide showed similar results, but their relevance to T-cell reactivity against native gp100 209-217 is uncertain. These results show that: (a) QS-21 and GM-CSF are superior to IFA as immunological adjuvants for vaccination against tyrosinase 370D peptide; and (b) with appropriate adjuvants, increased frequencies of peptide-specific T cells after vaccination can be detected by enzyme-linked immunospot without prolonged prestimulation in vitro.

Adult↗

Validation of an enzyme-linked immunosorbent assay for detection of mouse parvovirus infection in laboratory mice.

PURPOSE: Parvoviruses are among the most prevalent infectious agents in mouse colonies. Infection in laboratory mice is confirmed by detection of serum antibodies to these agents, and most diagnostic tests cannot distinguish serogroup of the infecting agent. The principal objective of the research reported here was to develop and validate a sensitive, serogroup-specific diagnostic test that will distinguish between mouse parvovirus (MPV) and minute virus of mice (MVM) infection. METHODS: The MPV VP2 protein was expressed in bacteria, purified by use of metal-chelation chromatography, and used as antigen in an ELISA. More than 580 sera from uninfected mice and experimentally or naturally infected mice were screened by MPV indirect fluorescent antibody (IFA) test, then were re-tested using the MPV ELISA to define test sensitivity and specificity. An additional 3,700 sera were screened using a variety of tests, including the MPV ELISA and recombinant NS1 ELISA (rNS1 ELISA). RESULTS: Using MPV IFA test results as a benchmark, the MPV ELISA had sensitivity of 92.3% and specificity of 99.8%. In addition, the MPV ELISA detected anti-viral antibodies at a higher dilution of serum than did the IFA test, and confirmed the infecting agent as MPV or MVM. When compared directly in a commercial laboratory, the MPV ELISA had higher sensitivity (90.3% versus 65%) than and similar specificity (98.3% versus 99.6%) to the rNS1 ELISA. CONCLUSION: The MPV VP2 ELISA provides a sensitive, serogroup-specific alternative for diagnosis and classification of parvovirus infection in laboratory mice.

Animals↗

Cat scratch disease. Survey on the presence of Bartonella henselae among cats of Tuscany.

To verify the presence of Bartonella henselae-infection in cats living in Tuscany (central Italy) serological and bacteriological surveys were carried out. The blood serum samples of 427 cats, 254 living in private houses and gardens and 173 in public or private catteries, were tested for anti-B. henselae antibodies by indirect immunofluorescence assay (IFA). Among these samples, 35 were examined by IFA to detect antibodies against Bartonella quintana. Bacteriological examinations were performed on the blood samples, collected in EDTA (ethylene diaminetetraacetic acid), of 18 cats (10 seropositive to B. henselae and 8 negative). From each of the same 18 specimens DNA was extracted and used as template in polymerase chain reaction (PCR). The primers p24E and p12B were employed in the PCR assay to amplify a 296 bp fragment of the Bartonella 16S rRNA gene. IFA detected 98 (22.95%) B. henselae-positive serum samples (40-40.82% from cats living in houses and gardens and 58-59.18% from cats of catteries) at different antibody titers (70 at 1:64 titer, 4 at 1:128, 22 at 1:256, 2 at 1:512). Among the 35 sera tested to detect antibodies against B. quintana, 9 (25.71%) resulted positive at 1:64 titer; all these samples showed higher antibody titers to B. henselae. Out of the 26 negative sera, 20 were negative to B. henselae too and 6 had antibodies against B. henselae at 1:64. Hemocultures gave negative results. PCR scored positive with DNA of 4 B. henselae-seropositive cats, two of which belonged to two children with cat scratch disease (CSD).

Animals↗

Development of monoclonal antibody based antigen capture ELISA to detect chikungunya virus antigen in mosquitoes.

BACKGROUND & OBJECTIVES: Chikungunya (CHIK) virus has caused numerous large outbreaks in India. No active or passive surveillance has been carried out since the last epidemic which occurred in 1971. For active surveillance, it is necessary to have a test, which can detect the virus from a large number of field-collected mosquitoes. METHODS: The present study describes the standardization of monoclonal antibody (MAb) based antigen capture ELISA to detect chikungunya virus antigen from the mosquitoes. CHIK virus antigen from suspension of experimentally infected mosquitoes and their progeny was captured on mouse polyclonal antibody, while biotinylated CHIK Mab was used as a probing antibody. CHIK virus antigen in the head squashes of virus inoculated mosquitoes was detected using indirect immunofluorescence antibody (IFA) test for confirmation of ELISA results. RESULTS: The ELISA test was sensitive enough to detect antigen even if a small fraction of a single infected mosquito homogenate was incorporated in the test. The IFA test failed to detect CHIK antigen in 10 and 25 microliters of suspension whereas with ELISA it was detected in all the samples. Progeny of Aedes aegypti and Ae. albopictus mosquitoes infected with chikungunya virus did not show the possibility of existence of transovarial transmission. INTERPRETATION & CONCLUSION: This test is rapid and simple since it can be completed in two days as compared to the conventional mosquito inoculation and IFA techniques, which require at least 10 days. There is an additional advantage with this test that a large number of samples can be processed, and the remaining homogenate of the mosquitoes can be used for screening other viruses. Experimental data raised using this test showed that transovarial transmission of this virus does not occur in these vector species.

Aedes↗

Comparative immunopotentiating properties of saponin and incomplete Freund's adjuvant coupled to affinity purified larval antigen of Hyalomma anatolicum anatolicum.

Comparative immunostimulatory properties of saponin and incomplete Freund's adjuvant (IFA) were studied using affinity purified 39 kDa larval antigen of Hyalomma anatolicum anatolicum. Significant antibody response to 39 kDa antigen was detected in the sera of rabbits immunized with both 39 kDa plus saponin and 39 kDa plus IFA in comparison to their corresponding control animals. Insignificant differences were noted in the antibody response between the animals of two immunized groups. Upon challenge, the animals immunized with 39 kDa plus IFA rejected 76.2 +/- 9.7% of larvae and 45.8 +/- 4.1% of adults while in group of animals injected with 39 kDa plus saponin rejected 80.9 +/- 11.2% of larvae and 47.2 +/- 5.7% of adults.

Animals↗

[Role of certain neurohumoral factors in development of pulmonary hypertonia in patients with interstitial lung diseases].

AIM: To ascertain the role of some neurohumoral factors--nitric oxide (NO), ACE, histamine--in development of pulmonary hypertension (PH) in patients with interstitial lung diseases (ILD). MATERIAL AND METHODS: A total of 32 ILD patients were examined. Of them 14 had idiopathic fibrosing alveolitis (IFA), 6 had exogenous allergic alveolitis (EAA) and 12 patients had ILD in diffuse disease of the connective tissue (ILD-DDCT). In addition to routine tests, those for NO, ACE, histamine, serotonin in plasma were performed; Doppler echocardiography and high-resolution computed tomography were conducted. In 9 patients the diagnosis was verified at thoracoscopic biopsy of the lung. The control group consisted of 16 healthy subjects. RESULTS: The highest mean pressure in the pulmonary artery (PmPA) was registered in IFA vs EAA and ILD-DDCT patients (p < 0.001). NO concentration in plasma was higher in ILD-DDCT than in control patients. In IFA and EAA the level of NO was like in controls. Concentration of NO in plasma of IDL patients correlated with high activity of the process. No correlation was found between ACE in plasma and PmPA, ACE levels were much higher in controls than in the ILD patients (p < 0.05). Histamine levels were higher in ILD patients than in controls being the highest in ILD-DDCT. Serotonin was insignificantly higher in ILD patients than in controls. CONCLUSION: Damage to the endothelium of the pulmonary arteries and imbalance of neurohumoral factors may be considered as a mechanism of development and stabilization of PH in ILD patients.

Adult↗

Evaluation of five commercial tests: complement fixation, microparticle agglutination, indirect immunofluorescence, enzyme-linked immunosorbent assay and latex agglutination, in comparison to immunoblotting for Mycoplasma pneumoniae serology.

A panel of 68 serum specimens from 41 subjects exhibiting various immunological patterns to Mycoplasma pneumoniae as determined by detection of a 180 kDa protein in immunoblotting was used to compare five commercially available tests based on different methods: complement fixation test (CFT), microparticle agglutination (MAG), indirect immunofluorescence assay (IFA), enzyme-linked immunosorbent assay (Elisa), and latex agglutination (LA). The tests were performed according to the manufacturers' instructions. For the determination of immunity to M pneumoniae, the five tests were in good accordance with immunoblotting: sensitivity was 100% for all the five assays, specificity ranged from 95.6% (MAG) to 82.6% (Elisa) and overall agreement ranged from 98.2% (MAG) to 92.8% (Elisa). The comparisons of antibody rates obtained by the four quantitative tests (CFT, MAG, IFA, Elisa) showed correlation coefficients ranging from 0.87 (CFT-IFA) to 0.67 (CFT-Elisa). Six significant antibody rises demonstrated by immunoblotting patterns were detected by all the tests but Elisa in one case. As a whole, the commercial assays gave satisfactory results for routine determination of immune status to M pneumoniae: CFT was the cheapest test and MAG and LA were the easiest to perform.

Adolescent↗

New look at chronic gastritis.

Chronic gastritis (CG) is classified as Type A (Autoimmune) and Type B [Bacterial-Helicobacter pylori (HP)]. Even Type A gastritis is initiated by HP (despite its low incidence in the gastric mucosa), as the incidence of antibodies against HP (anti-HP) in sera of patients with pernicious anemia (PA) is high. Anti-HP cross react with gastric mucosal antigens, including alpha and beta subunits of the proton pump (parietal cell antigen) and determine the damage in fundus-body and/or antral mucosa. Failure to develop intrinsic factor antibodies (IFA), determined by hereditary factors, explain the rarity of PA (despite wide prevalence of CG due to HP) in the Indian population. The necessity of separating severe chronic atrophic gastritis with histamine-fast achlorhydria from PA on the basis of absence or presence of IFA and restricting the diagnosis of PA only to those with IFA, cannot be overemphasized.

Adenosine Triphosphatases↗

Experimentally induced infection of cats with Borrelia burgdorferi.

To determine whether cats could be infected experimentally with Borrelia burgdorferi, 15 cats were inoculated with approximately 1,000 B burgdorferi. Seven cats were inoculated by the IV route, 2 by the oral route, 2 by the ocular route, and 4 by the oral-ocular route. Six control cats were inoculated with phosphate-buffered saline solution by the IV, oral, and ocular routes. Prior to the start of the study, all 21 cats were seronegative for B burgdorferi on the basis of results of the indirect fluorescent antibody (IFA) test, and their blood was B burgdorferi culture negative. All of the IV, orally, and ocularly inoculated cats developed IgG antibodies to B burgdorferi as detected by IFA testing. Of 4 oral-ocularly inoculated cats, 2 developed IFA-detectable antibodies and the remaining 2 cats developed low-titer response (1:128) on postinoculation (PI) day 10 only. All control cats remained seronegative. The organism was detected in blood smears from 2 of the IV inoculated cats on PI days 10 and 24 and from 2 oral-ocularly infected cats, 1 on PI days 17 and 24 and 1 on PI day 10. Spirochetes were not detected in the blood after PI day 24. The organism was isolated from tissues of only 1 cat (the lung of an ocularly inoculated cat necropsied at 7 months after inoculation). Spirochetes were not isolated from control cats. Neither clinical signs of infection nor gross or histologic abnormalities were found in any of the inoculated or control cats. Results indicate that cats are susceptible to infection with B burgdorferi, but clinically apparent disease may not be common.

Animals↗

Evaluation of a canine C6 ELISA Lyme disease test for the determination of the infection status of cats naturally exposed to Borrelia burgdorferi.

The efficacy of a commercially available in-office kit (SNAP 3Dx, IDEXX Laboratories) for detection of antibodies directed against an invariable region (IR6) of the B. burgdorferi surface protein VlsE (Vmp-like sequence, Expressed), a surface antigen of the spirochete recognized during active infection has been evaluated in dogs. The present study was conducted to determine whether this in-office test could be useful for detection of antibodies to B. burgdorferi in cats. Cats owned by clients of a veterinary hospital located in an area hyperendemic for Lyme disease were included in the study. When possible, cats with an outdoor lifestyle, bite wounds, or current tick infestation were recruited for the study to help ensure that animals with a likelihood of exposure to natural infection by B. burgdorferi would be included in the test group. Of the 24 cats tested, 17 samples were positive for antibodies to B. burgdorferi by the C6 ELISA kit. For all 17 of these samples, a duplicate sample tested by immunofluorescent assay (IFA) was in agreement with the ELISA. Five samples were negative by both assays. Two samples that were negative by the C6 ELISA test had low IFA titers (1:100). One of these two discrepant samples was negative and one was positive for antibodies to B. burgdorferi by the Western blot test. It was concluded that the C6 ELISA test performed with good agreement with the IFA and Western blot tests for detection of antibody to B. burgdorferi in the majority of cats tested. The test offers the advantages of producing a result rapidly (approximately 8 minutes), and it requires only two drops of serum, plasma, or whole blood.

Animals↗

[Tick borne zoonosis: selected clinical and diagnostic aspects].

Tick-borne zoonotic infections are among the most diffuse vector borne diseases: these large group of infections is caused by different microorganisms: Babesia spp., Borrelia spp., Rickettsia spp., Ehrlichia spp., Francisella tularensis, Coxiella burnetii) and tick-borne encephalitis virus. Babesiosis is caused by the protozoa (sporozoa) Babesia microti and it is quite rare in humans in Europe. The ixodids ticks are the competent vectors. A few symptomatic cases have been reported, mainly in splenectomized patients. The laboratory diagnosis is made by the microscopic identification of the parasites within the red blood cells in blood smears. The serologic diagnosis, based mainly upon IFA and WB techniques has only an epidemiological interest. Lyme borreliosis (Lyme disease) has been recognized as the most frequent vector borne disease in mild climate areas. The etiologic agent is a spirochete, belonging to the Borrelia burgdorferi sensu lato complex: B. burgdorferi sensu stricto, B. garinii and B. afzelii. Several additional species of this geno-complex have been identified but their pathogenic capability for humans still needs to be elucidated. Lyme borreliosis is clinically divided into three different clinical stages: the early disease, the disseminated infection and the persistent infection. Individual stages are caused by the diffusion of the spirochetes to different anatomic districts of the body. The main clinical symptoms are, for each stage: the erythema chronicum migrans in the early infection, the peripheral nerves and joint involvement in disseminated diseases and the acrodermatitis chronica atrophica (ACA) with central nervous system involvement in the late disseminated infection. The microbiological diagnosis is achieved by serologic techniques (IFA, EIA, WB) and by isolation of the spirochetes (in vitro culture and DNA amplification methods). Tick-borne relapsing fever (TBRF) is occasionally transmitted to humans by the soft ticks Ornithodorus and is caused by Borrelia spp. Different borreliae are responsible for TBRF in various geographic areas. The laboratory diagnosis is based upon the identification of spirochetes in peripheral blood by microscopic observation of Giemsa stained smears. Rickettsiosis diseases are caused worldwide by the obligate intracellular bacteria belonging to the genus Rickettsia. In the Mediterranean area the most frequently identified rickettsia is R. conorii, that causes the so called Mediterranean spotted fever. The serologic detection of a specific antibody response by IFA techniques is the most prominent tool for the diagnosis. In addition, the PCR method can be applied. Bacteria of the genus Ehrlichia are well known pathogens in veterinary medicine. Since the last decade their zoonotic capability has emerged and E. chafeensis, E. canis and the so called human granulocytic agent (HGE) have been identified in human diseases following a tick bite. The ehrlichiosis is characterized, in human, by a mild fever associated with lymphoadenopathy. The diagnosis is made on the identification of morulae (the intracytoplasmatic inclusion of the growing rickettsiae) in the white cells of peripheral blood. In addition the molecular diagnosis is also possible by PCR. Tick-borne encephalitis (TBE) is the only viral arthropod-borne encephalitis in Europe: it is caused by a flavivirus and it can also be transmitted by the ingestion of goat raw milk. The more relevant epidemiological figure is limited to the Alps, in particular to the Northern side (Austria). Isolated cases have been reported also in Italy. TBE is a benign self-limiting illness that usually recovers without any reliquate. The laboratory diagnosis is obtained by isolating the virus in cell cultures from the CSF or blood of acute phase patients. Serology is anyway the main laboratory tool to perform this diagnosis. Complement fixation and EIA IgM are the most used methods: the latter technique is particularly sensitive in early infection.

Animals↗

[Investigation of Arboviruses in Lancang river down-stream area in Yunnan province].

OBJECTIVE: To investigate the epidemic state of arboviruses in the downstream area of Lancang river in Yunnan province. METHODS: Mosquitoes were collected from Lancang river downstream area (including Lancang county and Simao city) during summer in 1998 and stored in liquid nitrogen after classification. The mosquito pools were homogenized and centrifuged, then the supernatant was inoculated into C6/36 cells for virus isolation. New isolates were identified by neutralization test(NT), ELISA, immunofluorescence assay(IFA) and polyacrylamid gel electrophoresis(PAGE). RESULTS: Totally 22 isolates of arbovirus were obtained from 233 mosquito pools by inoculation of C6/36 cells and positive rate of the isolation was 9.4%. Ten strains were resistant to both ether and 5 prime-IDU. So they were non-enveloped double-stranded (ds) RNA virus. Twelve segmented RNAs were shown by PAGE and PAGE profiles from the ten strains were 6-6 with minor variation. The isolates can be neutralized by immunized mouse ascites fluid of BJ95-75 strains of coltivirus by NT, and reacted with monoclonal antibody against BJ95-75 by ELISA. These viruses were identified as coltivirus. Nine isolates were sensitive to ether and resistant to 5 prime-IDU. So they were non-enveloped RNA viruses. PAGE showed 10 segmented RNA, and they were identified to be orbiviruses. Three isolations were sensitive to ether. One of them can be neutralized with JEV A2 strain antibody by NT and was positive to the homologous antibody by IFA. It was identified being strain of JE virus. One strain(YN92-4) can be reacted with anti-bunyavirus group specific immune ascites fluid by both IFA and ElISA, but reacted neither with anti-alpha virus group, nor with anti-flavivirus group JE virus ascites fluid. The virions are spherical and about 87 nm in diameter with surface projections by negative staining. Conclusion Twenty-two isolates have been obtained from wild caught-mosquitoes of Lancang river down-stream area in Yunnan province. Among them ten, nine, one and one were identified as coltivirus, orbivirus, JE virus and bunyavirus, respectively. One is under identification. This is the first report on bunyavirus isolated from mosquitoes in China.

Animals↗

Indirect fluorescent antibody test in diagnosis of acute fasciolitic syndrome.

250 patients with fever, hepatomegaly and eosinophilia were investigated for parasitic infection by stool, urine, thick blood smear examination and sero-diagnosis (ELISA for anti-Toxocara and IFA for anti-Fasciola). Those with patent parasitic infections (41) and anti-Toxocara seropositivity (10) were excluded from follow up. The other were 3 months followed up. The detected infections were 10 Ascaris, 15 Strongyloides, 20 Toxocara, and 25 Schistosoma mansoni. Other 30 were positive for Fasciola eggs. No other infections were found in the remained 109 patients. IFA anti-Fasciola antibody titres were significantly higher among Fasciola group than any other groups. It is concluded that IFA anti-Fasciola antibody titre greater than or equal to 1/256 with ethanol fixed antigen sections is of value in diagnosis of acute fasciolitic hepatic syndrome.

Acute Disease↗