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PubMed1987

Evaluation of quaternary aminoethyl-Sephadex A50 column chromatography for detection of anti-cytomegalovirus immunoglobulin M.

The sensitivity of an indirect fluorescent antibody (IFA) test (screening test) for the detection of antibodies to cytomegalovirus (CMV) was examined by using 128 serum specimens and quaternary aminoethyl (QAE)-Sephadex A50 column chromatography to separate IgM from IgG class antibodies. Of these 128 rheumatoid factor-negative serum samples, only 54 (42%) continued to produce fluorescence after the IgG was removed by the Sephadex column (yielding a theoretical 1:10 dilution) and the samples were retested for IgM (confirming test). Of the 54 specimens that were positive on both the screening and confirming IgM IFA assays, 53 (98%) had titers of 1:80 or more. Of the 74 sera that were negative on the confirming test, only 10 (14%) had titers of 1:80 on the initial screening test, and none was greater than 1:80. Immunoglobulin measurements of eight serum samples before and after column treatment indicated excellent IgG removal but poor IgM recovery (a mean loss of 85% of the total IgM). Comparison of the time needed to detect IgM in CMV culture-positive patients demonstrated an increase from 17.1 to 28.3 days (serum tested by IFA before and after column separation, respectively). Therefore, confirmation of anti-CMV IgM in serum specimens by QAE-Sephadex A50 column chromatography seems to be of low sensitivity.

Antibodies, Viral↗
PubMed2003

Recovery and detection of Cryptosporidium parvum oocysts from water samples using continuous flow centrifugation.

Continuous flow centrifugation (CFC) was used in conjunction with immunomagnetic separation (IMS) and immunofluorescence microscopy (IFA) and nested PCR to recover and detect oocysts of Cryptosporidium parvum and cysts of Giardia intestinalis from 10L volumes of source water samples. Using a spiking dose of 100 oocysts, nine of 10 runs were positive by IFA, with a mean recovery of 4.4+/-2.27 oocysts; when another 10 runs were analyzed using nested PCR to the TRAP C-1 and Cp41 genes, nine of 10 were positive with both PCR assays. When the spiking dose was reduced to 10 oocysts in 10L, 10 of 12 runs were positive by IFA, with a mean oocyst recovery of 3.25+/-3.25 oocysts. When 10 cysts of Giardia intestinalis were co-spiked with oocysts into 10L of source water, five of seven runs were positive, with a mean cyst recovery of x=0.85+/-0.7. When 10 oocysts (enumerated using a fluorescence activated cell sorter) were spiked into 10L volumes of tap water, one of 10 runs was positive, with one oocyst detected. For the majority of the source water samples, turbidities of the source water samples ranged from 1.1 to 22 NTU, but exceeded 100 NTU for some samples collected when sediment was disturbed. The turbidities of pellets recovered using CFC and resuspended in 10 mL of water were very high (exceeding 500 NTU for the source water-derived pellets and 100 NTU for the tap water-derived pellets). While not as efficient as existing capsule-filtration based methods (i.e., US EPA methods 1622/1623), CFC and IMS may provide a more rapid and economical alternative for isolation of C. parvum oocysts from highly turbid water samples containing small quantities of oocysts.

Animals↗
PubMed2002

Evaluation of an enzyme-linked immunosorbent assay for detection of antibodies to Junin virus in rodents.

Junin virus is the etiological agent of Argentine hemorrhagic fever, a serious rodent-borne disease. An enzyme-linked immunosorbent assay (ELISA) to detect Junin virus IgG antibodies in rodents was evaluated using sera from 27 Calomys musculinus and five Calomys laucha, inoculated experimentally with a live attenuated strain of this arenavirus. The test performance was compared against an indirect immunofluorescence assay (IFA). The ELISA had a sensitivity and specificity of 100% and a reproducibility of 87.9% for samples with titers above the selected cut-off value. IFA had lower sensitivity (53%) with the same specificity. The ELISA results were similar, whether carried out on whole blood or serum samples, thus eliminating the need for serum separation. A high correlation (K=0.86) between ELISA and IFA results was obtained from 1011 wild sigmodontine and murine rodents collected within and outside of the Argentine hemorrhagic fever endemic area. These results indicate that Junin virus IgG ELISA is the most suitable assay for detection of Junin virus antibodies in rodent samples.

Animals↗
PubMed1997

Detection of IgG antibody to Epstein-Barr virus viral capsid antigen in saliva by antibody capture radioimmunoassay.

A 'G' antibody capture radioimmunoassay (GACRIA) to detect IgG to Epstein-Barr virus (EBV) viral capsid antigen (VCA) in saliva is described. The monoclonal antibody to EBV VCA used in the GACRIA bound non-specifically when testing saliva samples having a total IgG content of less than 2.0 mg/l, so giving false positive results. This problem was overcome by including 0.5% EBV-negative human serum in the monoclonal antibody diluent. The performance of the assay was then evaluated by comparing the GACRIA test on serum and saliva samples to indirect immunofluorescence assay (IFA) results on sera using a panel of paired serum/saliva samples. Compared to the corresponding serum IFA the saliva GACRIA had a sensitivity and specificity of 93.5 and 100%, respectively. Although less sensitive than IFA on serum samples, the saliva GACRIA is sufficiently sensitive to be used for epidemiological screening and will enable testing for anti-EBV VCA to be carried out easily and on a wide scale.

Antibodies, Viral↗
PubMed1998

The muskrat (Ondatra zibethicus) as a new reservoir for puumala-like hantavirus strains in Europe.

We have used an indirect immunofluorescent assay (IFA) and reverse transcription-PCR (RT-PCR) to screen the sera and tissues of muskrats (Ondatra zibethica) caught in the northwest of Brandenburg and in the northeast of Saxony-Anhalt, Germany, for hantavirus infection. Kidney and/or lung tissue from 6 (3.1%, CI = 1.1-6.5%) out of 197 muskrats were found to be positive for genomic sequences of hantavirus by RT-PCR. We could also demonstrate that 14 (5%, CI = 2.9-8.7%) out of 266 muskrat's sera available for testing contained hantavirus-specific antibodies in IFA. Thus, a total of 8% of the investigated muskrat population was found to be positive for hantavirus infection by RT-PCR and IFA. None of the animals was found positive in both tests. Further analysis of the RT-PCR amplified fragments by genomic sequencing revealed sequences mostly related to the puumala (PUU) S segment sequence of the Hällnäs B1 hantavirus strain (97-99% similarity). Our data therefore demonstrate that Ondatra zibethicus serves as an additional reservoir for puumala-like hantavirus strains in Europe. The epidemiological implications of this finding for hantavirus infection in Europe and elsewhere are discussed.

Amino Acid Sequence↗
PubMed1998

Release of tetanus toxoid from adjuvants and PLGA microspheres: how experimental set-up and surface adsorption fool the pattern.

The classical adjuvants alum and Freund's Incomplete Adjuvant (IFA) are frequently used as references for the design of new adjuvants and antigen delivery systems, e.g., microspheres (MS). Poly(dl-lactic-co-glycolic acid) (PLGA) MS have been proposed for delivering antigen booster doses in vivo after a single injection. However, as antigen release kinetics from conventional adjuvants are generally unknown, it appears presumptuous to propose a desired antigen release pattern from PLGA MS. Therefore, we have studied the tetanus toxoid (Ttxd) in vitro release from alum, IFA formulations and MS in four different test systems. The results showed a stronger Ttxd association to alum than to IFA, and the release from both formulations lasted between 3-9 days. The total of ELISA-responsive antigen released was 60-85% of the actual dose. Both the total amount and the prolongation of release depended on the Ttxd dose. Furthermore, the incomplete in vitro release of Ttxd from the adjuvants and also from PLGA 50:50 MS was shown to be partly due to experimental conditions. Typically, Ttxd adsorbed on the glass vials used for the release test and also on the surface of the PLGA 50:50 MS, wherefrom it was released. In conclusion, the test system depending rate and quantity of release observed evidence the limitations of in vitro release data. Finally, for mimicking conventional vaccination schedules, i.e. injections typically at time points 0, 1, 3, and 12-24 months, PLGA MS should release antigen doses at the corresponding time points, and the release pulse should only last for a few days.

Adjuvants, Immunologic↗
PubMed2000

Comparison of two laboratory methods for the determination of serum resistance in Borrelia burgdorferi isolates.

A growth inhibition assay (GIA) and an immunofluorescence test detecting deposited complement components C6 and C9 were compared for their ability to classify Borrelia isolates with respect to their resistance to non-immune human serum (NHS). In both assays a total of 34 Borrelia isolates of all three human pathogenic genospecies were tested. Interestingly, 95% of the serum-sensitive or intermediate serum-sensitive isolates belonged to the genospecies B. burgdorferi s. s. and B. garinii, whereas most B. afzelii isolates (83%) proved serum-resistant. Consequently, a strong correlation between the assignment of the isolates to the different genospecies and their degree of serum sensitivity was seen. These findings were supported strongly by the quantitative analysis of the deposited complement components and the location of the terminal complement complex on the bacterial surface as detected by means of immunoelectron microscopy. The GIA displayed an obvious lack of sensitivity to slow growing isolates, whereas the IFA allowed classification of all Borrelia isolates. Discrimination between serum-sensitive and serum-resistant isolates in the IFA was the most specific provided that the detection of C6 and C9 was incorporated into the final classification of isolates. Accordingly, both assays, turned out to be effective and reliable tools for the investigation of borrelial serum sensitivity. The IFA, however, is regarded as superior to the GIA owing to the obvious ease of performance and its rapid capability for the classification of even very slow growing isolates.

Borrelia burgdorferi Group↗
PubMed1986

Experimental inoculation of dogs with Borrelia burgdorferi.

To determine if dogs could serve as a reservoir for Borrelia burgdorferi, three beagles were inoculated subcutaneously (SQ) with 200 laboratory cultured spirochetes which were originally isolated from blood of a Peromyscus leucopus from Ft. McCoy, Wisc. One four month old beagle was inoculated SQ with 5 ground Ixodes dammini from Shelter Island, N.Y. which came from an area with a 50% B. burgdorferi tick infection rate; and another uninfected four month old beagle was housed loose on the floor with the tick inoculated dog. All three spirochete inoculated beagles developed IFA antibody titers to B. burgdorferi of (7 log2) to (8 log2) by day 28 post inoculation. All were apparently healthy and no spirochetes were cultured from the blood. In an attempt to exacerbate the disease two of the dogs were given 3 mg of dexamethasone on day 68 post inoculation. B. burgdorferi was isolated from blood of all these dogs on days 4 and 97 days post inoculation. The tick inoculated dog developed a B. burgdorferi IFA antibody titer of (10 log2) by day 14 post inoculation. The contact exposed dog also developed a B. burgdorferi IFA antibody titer of (7 log2) on post contact day 21 indicating contact infection. B. burgdorferi was not isolated from either of these dogs. These results indicate that, contact transmission of B. burgdorferi may occur between dogs, dogs can be subclinically infected with B. burgdorferi and have persistent infections.

Animals↗
PubMed1998

Whole-killed gp120-depleted HIV-1 antigen in a murine model for prophylactic vaccination.

In this study, the effects were examined of dose and adjuvant of whole-killed gp120-depleted HIV-1 antigen on antibody and cytokine responses in a murine model. Immunization with increasing doses of inactivated HIV-1 antigen in Incomplete Freund's Adjuvant (IFA) resulted in increased production of IL-4 and IgG1 antibody with decreased production of interferon gamma. Immunization with inactivated HIV-1 antigen in Detox PC adjuvant produced TH1 type predominant cytokine patterns along with IgG2a subclass antibody. Higher levels of interferon gamma were associated with immunization with inactivated HIV-1 antigen in Detox PC compared with inactivated HIV-1 in IFA or inactivated HIV-1 in saline. Inactivated HIV-1 antigen in Detox PC adjuvant produced a trend of lower levels of the beta-chemokine MIP-1 alpha compared with inactivated HIV-1 in IFA or saline. Dose and adjuvant play an important role in the type of immune response elicited to a whole-killed HIV vaccine. Low doses of inactivated HIV-1 antigen in Detox PC adjuvant are currently being studied in animal models in order to optimize cell-mediated immunity against HIV infection.

AIDS Vaccines↗
PubMed2000

In vitro immune function after vaccination with an inactivated, gp120-depleted HIV-1 antigen with immunostimulatory oligodeoxynucleotides.

We examined the effect of a synthetic oligodeoxynucleotide containing unmethylated CpG immunostimulatory sequences (ISS) as an adjuvant for an HIV-1 immunogen (inactivated, gp120-depleted HIV-1 virus particles). The addition of the ISS to HIV-1 in incomplete Freund's adjuvant (IFA) was the optimal combination for the production of HIV-1-specific immune responses as measured by IFN-gamma (p=0.002) and IgG antibody production. Furthermore, the group that was immunized with HIV/IFA/ISS, as well as the group that received HIV-1 in complete Freund's adjuvant (CFA), stimulated significantly more RANTES (a beta-chemokine) (p=0.002) production from lymph node cells. These results suggest that the addition of CpG immunostimulatory sequences to HIV antigens in IFA may optimize HIV-1-specific immune responses and provides further rationale for the testing of ISS in combination with gp120-depleted whole-killed HIV-1 in IFA as a potential preventive or therapeutic HIV-1 vaccine.

AIDS Vaccines↗
PubMed2000

Intermediate filament proteins in echinoderm coelomocytes.

The presence and organization of intermediate filament (IF) proteins in petaloid coelomocytes from two species of echinoderms, the sea urchin Strongylocentrotus droehachiensis and the sea cucumber Cucumaria frondosa, were studied. Two monoclonal antibodies (IFA and Ah6) and one polyclonal antibody (W3-1) that together recognize invertebrate as well as vertebrate IF proteins were used to probe coelomocytes by immunofluorescence and immunoblotting methods. All three antibodies cross-reacted with a single Mr 68,000 sea urchin lamin, as well as two putative lamin isoforms of approximately Mr 70,000 and 68,000 in sea cucumber coelomocytes. Both IFA and Ah6 labeled granular material in the cytoplasm of sea urchin coelomocytes; by contrast, IFA labeling revealed a striking network of reticular material irregularly arrayed within the central regions of the sea cucumber coelomocyte cytoplasm. In addition, foci of Ah6-positive material were present in coelomocyte nuclei from both species. Comparison of immunoblotting patterns among whole cell and isolated nuclear preparations suggest that the cytoplasmic IF-like material is composed of Mr 46,000 and 58,000 polypeptides, while Mr 215,000 and 185,000 proteins are candidates for the immunoreactive nuclear foci. Further study of the functions of these non-filamentous arrays of IF proteins may furnish valuable insights into the evolution of IF function within vertebrate cells, particularly with respect to certain cytoplasmic and nuclear regulatory functions with which IF proteins have been speculated to be involved.

Amino Acid Sequence↗
PubMed1976

[Trial serologic detection of Plasmodium carriers among blood donors of the Centre de Transfusion Sanguine de Marseiile].

This paper is a survey of possible reservoir hosts of Plasmodium among donors of the blood transfusion Center of Marseille, by IFA test with P. falciparum and P. cynomolgi bastianellii antigens. It was found 15,7 per cent of positivity among 644 sera carefully selected during 6 months. Nord Africa and West Africa were the most dangerous countries, the former because of the great number of donors (332 of 744), the later because of the high rate (15,8 per cent) of positivity. The incidence of the say (frequency and long time) in infested countries and the interval from tate (15,8 per cent) of positivity. The incidence of the stay (frequency and long time) in infested countries and the interval from the last infestation possibility to the IFA test are very important factors. A prevention scheme is proposed: The donors staying during a very long time in infested countries will generally be used only for preparing plasma fractions by ethanol precipitations. For the others donors the blood will be used for preparing whole blood, red cells, fresh or frozen plasma, platelets and leucocytes preparation, in which a few parasites can be present, inly if a IFA test is negative 4 months after their coming back in our country.

Africa, Northern↗
PubMed1981

[Post-transfusion malaria. Current state of the problem].

A global review of the problem of malaria accidentally transmitted by blood transfusion is reported in France during these last twenty years. A survey by IFA test of possible reservoir hosts of Plasmodium among 2571 donors from the blood transfusion Centers of Marseille and Nice is reported. The incidence of the stay (frequency and longtime) in infested countries and the interval from the last infestation possible to the IFA test are very important factors. Several preventive methods are discussed. The author proposes this scheme: the donors frequently going to infested countries will be used only for preparing chemical plasma fractions. For the other donors the blood will be used for preparing whole blood, red cells, fresh or frozen plasma, platelet and leucocyte preparations in which a few parasites can be present, only if IFA test is negative 4 months after their coming back in our country. Nevertheless this screening beginning by an interrogation can be fallible. Therefore, there is no absolute preventive method. Physicians, Surgeons and Anesthetists must be forewarned.

Blood Donors↗
PubMed1985

[Screening for Chagas disease by immunoenzymology: comparison of ELISA with immunofluorescence and indirect hemagglutination in 976 blood donors].

The efficiency of an immunoenzymatic technique (ELISA) for the systematic research of Chagas' disease in blood donors was compared with one of 2 well-known methods, indirect haemagglutination (IHA) and indirect fluoro immuno assay (IFA). For the ELISA technique two different antigenic extracts from epimastigote culture forms of T. cruzi, were used for sensitizing the polystyrene plates: a crude extract (Ag R) and a delipidized one (Ag B). Firstly the authors tested these 3 techniques in 5 control groups: sera from Chagas' disease, negative control sera, sera from visceral leishmaniasis, african trypanosomiasis and finally monoclonal gammapathies, the high levels of blood proteins being a possible cause of false positives. Secondly the screening of Chagas' disease was performed in the same way in 976 blood donors from Recife, Brazil. In the case of the Ag-R extract used in the ELISA technique a high cross-reactivity was found with visceral leishmaniasis sera, along a risk of false positives with gammapathic ones. The sensitivity of this technique was found to be high (3,3 +/- 1 p. cent of positive blood donors) and a very good correlation was found with the reference techniques, IFA and IHA, the sensitivity of which is lower (2,3 +/- 1 and 1,7 +/- p. cent). The use of a delipidized antigenic extract (Ag B) for the ELISA technique is not suitable, in spite of an apparent higher specificity: indeed, the positives rate is high (11,5 +/- 0,2 p. cent), but the correlation is very weak or non existent in the case of IHA or IFA. In conclusion, the ELISA technique using a crude extract of T. cruzi appears to be a very convenient method for screening blood donors with Chagas' disease, the lack of specificity due to leishmaniasis or monoclonal blood proteins not posing any real problem to blood banking.

Blood Donors↗
PubMed1998

Experimental infection of cattle with several Borrelia burgdorferi sensu lato strains; immunological heterogeneity of strains as revealed in serological tests.

Twenty-three experimental cattle, mainly calves, were each inoculated 1-3 times with one of ten Finnish Borrelia burgdorferi sensu lato strains. All three genospecies were represented. Borreliae were administered mainly by both intravenous (about 10(6) to 10(9) spirochaetes) and intradermal (10(4)) routes, and on six occasions subcutaneously (10(3)) only. For infectivity control and comparison purposes mice and rabbits were inoculated simultaneously. Immune responses in cattle were monitored both with whole-cell sonicate enzyme-linked immunosorbent assay (IgG-ELISA) and indirect immunofluorescent assay (IgM-IgG-IFA). Five Finnish strains and the American strain B31 were used as antigens. No clinical signs of borreliosis were observed. Of the strains, 7/10 were interpreted by the immune responses to have caused relatively short-term subclinical infections of varying intensity. Borreliae could not be isolated from blood or other organ specimens of cattle. A rough estimate of the mean infectious dose in the conditions of experiments is 10(6) to 10(7) organisms. In conclusion, the overall result appears to argue a low susceptibility of cattle to clinical borreliosis, at least when infected by Finnish strains of the agent. Significant antigen-specific differences were observed both by ELISA and IFA in detection and quantification of immune responses. As a rule, the homologous antigen was found to be the most sensitive. Genospecies differences were mostly distinct. Antigens of two Borrelia garinii isolates proved practically equal in sensitivity, whereas major differences were displayed between two Borrelia afzelii antigens. In an IFA study, an American (B31) and a Finnish B. burgdorferi sensu stricto strain proved equally sensitive as antigens. In two relatively strong primary immune responses the antigen-specific measurement differences were such that diagnostically in a cross-sectional study only the homologous antigen or an antigen of the same genospecies would have been sufficiently sensitive to show a positive result.

Animals↗
PubMed1997

Significance of leukocyte concentration in the performance of the quantitative cytomegalovirus (CMV) antigenemia assay.

BACKGROUND: The quantitative cytomegalovirus-antigenemia (CMV-Ag) assay is an important technology in the regimen of tests utilized in the care and management of acquired immunodeficiency syndrome (AIDS) and other immunocompromised patient groups. Performance of this assay is contingent upon the appropriate processing of the polymorphonuclear leukocyte (PMNL) compartment of the peripheral blood. However, a cell input standard in the performance of the CMV-Ag assay, has not been established. Interpretive differences between laboratories utilizing the CMV-Ag assay may reflect this lack of test uniformity. OBJECTIVES: To determine the effect of different PMNL concentrations on the quantitation of CMV in peripheral blood. The leukocyte concentration resulting in optimal rates of viral detection, will be compared with the shell vial assay-indirect immunofluorescent assay (SVA-IFA), and conventional tube culture (TC-CPE). STUDY DESIGN: A total of 74 freshly collected blood specimens were tested by the CMV-Ag assay, using cytospin preparations consisting of 2 x 10(5), 4 x 10(5) and in preliminary experiments, 8 x 10(5) PMNLs/slide. Data obtained from these studies were compared to SVA-IFA and TC-CPE. Viral load was monitored among 11 symptomatic patients through sequential testing of these patients at the start of ganciclovir (GCV), foscarnet (PFA), or combination drug therapy. RESULTS: Among 74 blood specimens tested by the CMV-Ag assay, cytospin preparations consisting of 4 x 10(5) compared with 2 x 10(5) PMNLs/slide, affected a mean positive cell increase of 215% (P = 0.03). PMNL slide preparations consisting 8 x 10(5) cells produced background levels which prevented accurate reading of slides. The CMV-Ag assay was more sensitive than the SVA-IFA, but equivalent to TC-CPE. Among 11 patients started on drug therapy, viral load was markedly reduced in 8 within 2-3 weeks; three patients (2 deceased within 3 weeks after receiving therapy), showed no decrease in viral load. One patient was identified as harboring a PFA resistant strain. CONCLUSIONS: A PMNL concentration of 4 x 10(5) cells facilitated the reading of CMV-Ag assay slide preparations. The modified CMV-Ag assay furthermore, is applicable in the monitoring of viral load for the tracking of susceptible or resistant CMV strains.

Antigens, Viral↗
PubMed1997

Detection of respiratory syncytial virus, parainfluenzavirus 3, adenovirus and rhinovirus sequences in respiratory tract of infants by polymerase chain reaction and hybridization.

BACKGROUND: Immunofluorescence assay (IFA) of viral antigens in nasal aspirates is largely used for the diagnosis of respiratory syncytial virus (RSV), parainfluenzavirus (PIV) type 3 and adenovirus (AdV) infections, whilst rhinovirus (RV) are detected by virus isolation technique (VIT) only. Using the two techniques, IFA and VIT, a significant number of specimens remain negative in spite of clinical and epidemiological presumptions of viral infection. OBJECTIVES AND STUDY DESIGN: The polymerase chain reaction (PCR) should improve the sensitivity of viral detection in clinical specimens. From October 1995 to March 1996, 277 nasal aspirates from hospitalized infants were tested simultaneously by IFA, VIT, polymerase chain reaction and hybridization with a DNA enzyme immunoassay (PCR-EIA) for RSV, PIV-3, AdV and RV. RESULTS: RSV were detected in 177 (64%) samples, PIV-3 in 23 (8%), RV in 40 (14%), and AdV in 30 (10%). PCR-EIA detected RSV in more samples 173 (62%) than IFA/VIT: 109 (39%) (P < 10(-7)). In most cases (79%), RSV-infected infants had lower respiratory tract disease, and routine and PCR techniques were positive. Out of the 23 PIV-3 infections, 12 were IFA/VIT- and PCR-EIA-positive, and 11 IFA/VIT-negative and PCR-EIA-positive. For RV, 35 (87%) specimens were PCR EIA-positive and 11 (27%) culture-positive; for AdV 30 samples were PCR-EIA-positive and four were culture-positive. Simultaneous viral infections were revealed in a significantly higher proportion than in conventional techniques: 18% (50/277) versus 2.5% (7/277); P < 10(-7). One RSV infection in four was associated with the presence of another virus, mainly PIV-3 (16 cases) and AdV (13 cases). CONCLUSIONS: PCR-EIA detects more positive-specimens than IFA/VIT, 1.5 times more for RSV, 1.9 for PIV-3, 4 for RV and 10 for AdV, respectively. This increased sensitivity of viral detection by PCR-EIA compared to the IFA/VIT could suggest that samples containing low levels of virus are missed by routine methods IFA/VIT, and consequently, RSV or PIV-3, and above all RV or AdV are overlooked as agents of respiratory diseases. However, apart from the fact that the economic and convenient aspects of virus diagnostic cannot be missed, it is difficult to answer the following questions: what is the meaning of the detection of a viral sequences in nasal aspirates of infants, or may PCR have detected virus in patients who would not developed disease?

Adenoviridae↗
PubMed2003

Immunization with myelin or recombinant Nogo-66/MAG in alum promotes axon regeneration and sprouting after corticospinal tract lesions in the spinal cord.

We have shown previously that immunization with myelin in incomplete Freund's adjuvant (IFA) is able to promote robust regeneration of corticospinal tract fibers in adult mice. In the present study the effectiveness of such immunization with myelin was compared to that of a combination of two axon growth inhibitors in myelin, Nogo-66 (the 66-amino-acid inhibitory region of Nogo-A) and myelin-associated glycoprotein (MAG). The effectiveness of two adjuvants, IFA and aluminum hydroxide (Alum), was also compared, the latter being one that can be used in humans. In addition, larger dorsal overhemisections were made at the lower thoracic level, which resulted in a larger scar. These studies were carried out in SJL/J mice, a mouse strain that is susceptible to autoimmune experimental allergic encephalomyelitis (EAE). None of the immunized mice developed EAE. Long-distance axon regeneration and sprouting of the corticospinal tract was seen in myelin and Nogo-66/MAG immunized mice. Alum was as effective or better than IFA as the adjuvant. Overall, the robustness of axon growth and sprouting was greater in mice immunized with myelin. The abundance of this growth was less than in our earlier work in which smaller lesions were made, pointing to the possible influence of inhibitors in the scar. This work shows, however, that axon growth inhibitors in myelin can be selectively blocked using this immunization approach to promote long-distance axon regeneration in the spinal cord.

Aluminum Hydroxide↗