Strong Dependence of Atmospheric Feedbacks on Mixed-Phase Microphysics and Aerosol-Cloud Interactions in HadGEM3.

We analyze the atmospheric processes that explain the large changes in radiative feedbacks between the two latest climate configurations of the Hadley Centre Global Environmental model. We use a large set of atmosphere-only climate change simulations (amip and amip-p4K) to separate the contributions to the differences in feedback parameter from all the atmospheric model developments between the two latest model configurations. We show that the differences are mostly driven by changes in the shortwave cloud radiative feedback in the midlatitudes, mainly over the Southern Ocean. Two new schemes explain most of the differences: the introduction of a new aerosol scheme and the development of a new mixed-phase cloud scheme. Both schemes reduce the strength of the preexisting shortwave negative cloud feedback in the midlatitudes. The new aerosol scheme dampens a strong aerosol-cloud interaction, and it also suppresses a negative clear-sky shortwave feedback. The mixed-phase scheme increases the amount of cloud liquid water path (LWP) in the present day and reduces the increase in LWP with warming. Both changes contribute to reducing the negative radiative feedback of the increase of LWP in the warmer climate. The mixed-phase scheme also enhances a strong, preexisting, positive cloud fraction feedback. We assess the realism of the changes by comparing present-day simulations against observations and discuss avenues that could help constrain the relevant processes.

Theory of growth by differential sedimentation, with application to snowflake formation.

A simple model of irreversible aggregation under differential sedimentation of particles in a fluid is presented. The structure of the aggregates produced by this process is found to feed back on the dynamics in such a way as to stabilize both the exponents controlling the growth rate, and the fractal dimension of the clusters produced at readily predictable values. The aggregation of ice crystals to form snowflakes is considered as a potential application of the model.

Comparison of a commercial enzyme-linked immunosorbent assay with immunofluorescence and complement fixation tests for detection of Coxiella burnetii (Q fever) immunoglobulin M.

A commercially available enzyme-linked immunosorbent assay (ELISA) for the diagnosis of Q fever (PanBio Coxiella burnetii immunoglobulin M [IgM] ELISA, QFM-200) was compared to the indirect fluorescent antibody test (IFAT) for C. burnetii IgM and the complement fixation test (CFT). The ELISA demonstrated 92% agreement with the reference method (IFAT), and gave a sensitivity of 99% (69 of 70 samples) and a specificity of 88% (106 of 121). Specificity can be increased with confirmation by IFAT. CFT was found to have a specificity of 90% (107 of 119), although it was lacking in sensitivity (73%; 51 of 70). No cross-reactivity was observed in the ELISA with serum samples from patients with mycoplasma (n = 6), chlamydia (n = 5), or legionella (n = 4) infections, although 2 of 5 patients with leptospirosis and 1 of 4 samples containing rheumatoid factor (RF) demonstrated positive results in the ELISA. Results indicate that the performance of the PanBio C. burnetii (Q fever) IgM ELISA (F = 187) is superior to that of CFT (F = 163), and consequently the ELISA should be a useful aid in the diagnosis of acute Q fever.

Difficulties with the serologic diagnosis of infectious mononucleosis: a review of the RCPA quality assurance programs.

The Royal College of Pathologists of Australasia's Quality Assurance Programs (QAP) on serologic assays for infectious Mononucleosis (IM) have identified a number of important problems in laboratory diagnosis of this condition. A wide range of assays for the diagnosis of acute Epstein-Barr virus (EBV) infection are available, although heterophile antibody tests are still the most frequently performed procedures for the diagnosis of IM. In 1993, eighty-six (54%) of the 159 participating laboratories performed only a heterophile test; of these, 71% did not provide an interpretation of their results and none mentioned the need for confirmatory EBV-specific antibody testing. This revealed a lack of appreciation that heterophile tests should only be used for screening, due to their inferior sensitivity of less than 50% in children and 80-90% in adults and specificity of 95% compared to EBV-specific assays. For these reasons the use of heterophile tests is discouraged. Although EBV-specific serology has traditionally been done by immunofluorescence (IF), the use of reliable ELISA methods using purified EBV-antigens is increasing. False negative EBV VCA IgM ELISA test results were obtained by laboratories using unpurified or unspecified VCA antigens. As only five laboratories used these tests in the 1992 QAP, the tests' true performances could not be properly assessed, suggesting the need for independent studies. Variable results were obtained by laboratories using commercial EBNA IgG assays (in addition to EBV IgM) suggesting that an assessment of these kits would also be worthwhile. The QAP reaffirm that reliable EBV IgM detection is the most useful test for the diagnosis of IM, even where other EBV markers are also used. In the 1992 and 1993 QAP, laboratory performance of both heterophile and EBV-specific tests was better with non-reactive serum specimens (99% (correct) than with reactive serum specimens (90% correct).

Herpes simplex virus type 2 infection of heterosexual men attending a sexual health centre.

OBJECTIVE: To identify risk factors, particularly circumcision status, associated with serological evidence of herpes simplex virus type 2 (HSV-2) infection of heterosexual men. DESIGN: A cross-sectional case-control study employing an anonymous delinked interviewer-administered questionnaire, clinical examination, and a type-specific serological test for HSV-2. PARTICIPANTS AND SETTING: Three hundred consecutive heterosexual male patients at a public sexually transmissible diseases (STD) clinic in Sydney, Australia. MAIN OUTCOME MEASURES: Associations between serological evidence of HSV-2 infection and history of genital herpes or contact with genital herpes, history of other common STDs, and demographic and behavioural factors such as age, education level, number of sexual partners and lack of circumcision. RESULTS: One hundred and ninety-four patients (64.7%) had antibodies to HSV-2 but only 24% of these gave a history of genital herpes. A history of genital herpes or sexual contact with genital herpes, reported total lifetime number of sexual partners, failure to complete high school and a history of non-gonococcal urethritis or genital warts were associated with serological evidence of HSV-2 infection at the univariate level. Neither increasing age nor lack of circumcision was associated with HSV-2 infection. Following multivariate analysis only the lifetime number of partners and failure to finish high school were significantly strong predictors of HSV-2 infection. CONCLUSION: This is the highest prevalence of HSV-2 infection ever detected in an Australian population and one of the highest recorded globally. As younger men were as commonly infected as older men, and an earlier (1985) study involving the same clinic yielded a lower prevalence, it appears that a high level of ongoing HSV-2 transmission is occurring among Sydney heterosexuals. Increased awareness of this fact could enhance safer sex campaigns.

Detection of immunoglobulin M antibodies to glycoprotein G-2 by western blot (immunoblot) for diagnosis of initial herpes simplex virus type 2 genital infections.

Western blots (immunoblots) for the detection of immunoglobulin M (IgM) antibodies specific for herpes simplex virus type 1 (HSV-1) and HSV-2 in patients' sera were developed. The locations of the type-specific glycoprotein G (gpG-2) of HSV-2 (92- and 140-kDa forms) and glycoprotein C of HSV-1 (gpC-1), which carries mostly type-specific antigenic epitopes, were checked with specific monoclonal antibodies. Western blot assays for IgM antibody to gpC-1 or gpG-2 were performed after depletion of IgG by precipitation with anti-human IgG. In patients with primary HSV-2 genital infections, seroconversion of IgM and IgG antibodies to both the 92- and 140-kDa forms of gpG-2 was observed, although both antibodies appeared in convalescent-phase serum after the first week. IgM and IgG antibodies to low-molecular-size polypeptides (40 to 65 kDa) were the first antibodies observed in patients with primary infection, but these antibodies were cross-reactive with HSV-1 and HSV-2. However, in patients with recurrent HSV-2 infections, IgG antibodies to both forms of gpG-2 and the low-molecular-size polypeptides were found no matter how early after onset the patient was bled, and IgM to gpG-2 did not appear. In patients with nonprimary initial genital HSV-2 infections, IgG antibody to HSV-1 was demonstrated in the first serum specimen, and HSV-2-specific IgM was found in 39% of the serum specimens. Hence, the Western blot assay can be used to test for IgM antibody to gpG-2, allowing for the retrospective diagnosis of inital HSV-2 infections and its use as a supplementary test to the gpG-2 IgG enzyme-linked immunosorbent assays developed elsewhere. In contrast, IgM antibody to gpG-2 is not usually detected in patients with recurrent HSV-2 infections.

The reliability of serological tests for the diagnosis of genital herpes: a critique.

The reliability and limitations of the currently used routine tests for herpes simplex virus type 2 (HSV-2) antibody in Australia are reviewed. Six case reports illustrate the clinical dangers of overinterpretation of the currently available kits and the need for a readily available specific HSV-2 antibody test. In Sydney, HSV-2 causes approximately 85% of primary genital herpes and > 95% of recurrent genital herpes. Due to the extensive serological cross-reactivity between HSV-1 and HSV-2, currently available "type specific" commercial assays cannot reliably distinguish between the 2. Isolation of herpes simplex virus (HSV) or detection of HSV antigen in vesicle fluid is the preferred diagnostic test but may be overlooked or patients may have no visible lesions. The only accurate techniques for detecting HSV-2 specific antibody are the Western blot assay and an enzymatic immunoassay using glycoprotein G (gG-2), a component of the HSV-2 envelope. These tests, which still are restricted to research laboratories can be used to accurately identify people with previous exposure to HSV-2 (IgG) or to diagnose primary infection where virus isolation has not been performed or is impossible. Current commercially available antibody tests may have extensive cross-reactivity.

Herpes simplex virus type 2 antibody in patients attending antenatal or STD clinics.

OBJECTIVE: To determine the prevalence of antibody to herpes simplex virus type 2 (HSV-2) in patients attending a general public antenatal clinic and three public sexually transmitted disease (STD) clinics in Sydney. BACKGROUND: Highly specific tests for herpes simplex type 2 antibody, using the glycoprotein G2, have been recently introduced, allowing determination of past asymptomatic infection. Overseas studies have confirmed the long held suspicion that asymptomatic infection is more common than clinical genital herpes. The seroprevalence of HSV-2 in antenatal and STD clinic patients varies markedly in different countries. These are the first data available for Australia by means of this highly specific test. DESIGN: Cross-sectional study of seroprevalence in these two patient groups. Sera used in the antenatal study were those submitted for routine antenatal screening for viral markers. PARTICIPANTS: Two hundred and twenty-nine consecutive patients attending the Westmead Hospital antenatal clinics, and 107 consecutive patients attending three public STD clinics. HYPOTHESES: That Australian populations show a relatively high prevalence of past asymptomatic infection with HSV-2; and that higher rates of infection will be found in patients attending STD clinics and with past or current histories of STDs. MAIN OUTCOME MEASURES: Comparison of HSV-2 seroprevalence between antenatal clinic patients and STD clinic patients; and associations of HSV-2 antibody with age, sex, occupation, country of birth, a history of current or past STDs and antibody to HSV-1. RESULTS: Antibody to HSV-2 was found in 14.5% of antenatal clinic patients and 40% of STD clinic patients. None of the antenatal patients and less than half of the seropositive STD clinic patients reported clinical genital herpes. Associations with age, socioeconomic status and previous HSV-1 infection were less marked than in studies from the United States. Female STD clinic patients had a significantly higher seroprevalence than males and three times the seroprevalence of age-matched antenatal clinic patients. The correlation between HSV-2 antibody and current gonorrhoea was more marked than that between HSV-2 and other STDs. CONCLUSION: Asymptomatic infection with HSV-2 is quite common in Australian antenatal patients and more common in patients with STDs, who have higher rates of sexual exposure.

The diagnosis of recent herpes simplex virus type 2 genital infections by the simplex-2 test.

The prevalence of complement-fixing (CF) antibody against the AG-4 early antigen of herpes simplex virus (HSV) type 2 (HSV-2) was determined in patients with culture confirmed HSV-2 genital herpes and control groups using a commercial HSV-2 early antigen (Simplex-2; Gene Link Australia Ltd). Eighty seven per cent of 39 sera collected between 14 and 28 days after confirmed primary and recurrent HSV-2 infection were positive. In acute sera collected between 2-10 days after onset the Simplex-2 test was negative in all 90 patients with presumed primary infection but positive in 53% of 230 sera from recurrent infection. A specificity of 90-94.5% was obtained by testing 36 patients with recent proven HSV-1 infection and 331 control group patients. The Simplex-2 test may be useful in some cases of culture-negative, clinically suspected genital HSV-2 lesions only when sera are collected between 14-28 days after primary and recurrent infection. Its lack of specificity makes it unsuitable for the routine diagnosis of recent HSV-2 infection in the general population.

Indirect ELISA for the detection of HSV-2 specific IgG and IgM antibodies with glycoprotein G (gG-2).

The glycoprotein G (gG-2) purified from HSV-2 infected cells has been reported to be useful for determination of HSV-2 type-specific antibodies using conventional ELISA formats. This study further confirmed the specificity of gG-2 and demonstrated the feasibility of a specific IgM assay. The gG-2 ELISA was developed to detect HSV-2 specific IgG and IgM antibodies in human sera with high levels of sensitivity and specificity. Of 45 patients with culture-proven recurrent HSV-2 genital infection 44 were reactive for gG-2 IgG. Of 30 sera from patients with culture-proven recent initial HSV-2 genital infection 29 were positive for gG-2 IgM. Three patients with primary HSV-2 genital infection showed gG-2 IgM in the convalescent but not in the acute sera. The IgG- and IgM-gG-2 ELISA showed high specificity. None of 40 sera from children were reactive by either assay. Only one of 94 sera from patients with antibody to herpesviruses other than HSV reacted in the IgG assay but none reacted in the IgM assay. There was no cross-reaction with sera from patients with proven HSV-1 infection with the gG-2 antigen. The results suggest that the IgG assay can be used for demonstration of past HSV-2 infection and the IgM assay for the diagnosis of HSV-2 in neonatal herpes and primary genital herpes, when cultures or rapid diagnostic techniques are unavailable.

Rapid diagnosis of acute Epstein-Barr virus infection by an indirect enzyme-linked immunosorbent assay for specific immunoglobulin M (IgM) antibody without rheumatoid factor and specific IgG interference.

An indirect enzyme-linked immunosorbent assay (ELISA) for detection of Epstein-Barr virus-specific immunoglobulin M (IgM) antibody was developed with commercial reagents. Sera containing rheumatoid factor (RF) (as little as 0.5 IU/ml) coupled with specific IgG resulted in false-positives in the ELISA. This interference was eliminated by the use of anti-human IgG antibodies to remove RF and IgG. Thus, pathogen-specific IgG complexes to which IgM-RF could be bound during the subsequent test were inhibited, and competition between specific IgG and IgM was also prevented. Of the 1,672 serum specimens tested, 353 were found to be Epstein-Barr virus IgM antibody positive by indirect immunofluorescence (IF). Compared with the IF test, the ELISA showed 96.6% sensitivity, 99.7% specificity, and 99% accuracy. Further evidence indicated that most of the 12 ELISA false-negatives were IF false-positives. There was a linear correlation between mean ELISA values and increasing IF titers (r = 0.96). However, the IF test has the disadvantages that it lacks automated reading and requires considerable technical expertise, both of which restrict the range of laboratories performing the test. The indirect ELISA has the advantages that it is simple and rapid and can be automated. All the reagents used in this assay are commercially available, have been prestandardized, and are stable.

Evaluation of rubella immune status by three commercial enzyme-linked immunosorbent assays.

Three commercial indirect enzyme-linked immunosorbent assays (ELISAs) (Enzygnost-Rubella, RUBELISA, and ORTHO Rubella) were evaluated for the determination of immune status by testing 1,090 serum specimens, 410 of which were from nonimmune patients. In comparison with the standard reference technique, the hemagglutination inhibition (HAI) test, the sensitivities of ORTHO Rubella (100%) and Enzygnost-Rubella (99.26%) were excellent, whereas the sensitivity of RUBELISA (95.60%) was marginally lower because of the inability of this assay to detect antibody in 22% of the serum specimens with HAI titers of 10 and 11% of sera with HAI titers of 20. The specificity of all three systems was greater than 97%. There was a linear correlation between mean ELISA values and increasing HAI titers (r greater than or equal to 0.94). Both ORTHO Rubella and Enzygnost-Rubella were shown to be suitable replacements for the HAI test, provided that an equivocal zone is incorporated in the ORTHO system and only unheated sera are used in the Enzygnost system.

Immunoglobulin responses to echovirus type 11 by enzyme linked immunosorbent assay: single-serum diagnosis of acute infection by specific IgM antibody.

An indirect solid-phase enzyme-linked immunosorbent assay (ELISA) for the determination of specific IgM and IgG antibodies to echovirus type 11 in a single dilution of serum was developed using partially purified echovirus type 11 bound to microplates. Whole serum was used for IgG antibody but prior to assaying for IgM antibody interfering IgG was removed by ion exchange chromatography. The ELISA for echovirus type 11 IgG antibody was a more sensitive, rapid, technically easier and less costly alternative to the neutralisation test. With the IgG ELISA 12 of 132 sera (10.6%) known to contain enterovirus antibodies other than echovirus type 11 were positive but it could not be determined to what extent this was due to the greater sensitivity of the ELISA or cross-reactions. The IgM ELISA was even more sensitive than the IgG ELISA with acute sera, and showed a reactivity in 4 of 36 sera (11.1%) with no detectable echovirus type 11 neutralising antibodies. Echovirus type 11 IgM antibody was detected in all sera collected after the first week of infection and up to 30 days after infection. However, it was only detected in 58% of sera collected during the first week after onset thus limiting its use for rapid diagnosis. The echovirus type 11 IgM ELISA appears to have considerable laboratory diagnostic potential when a rising antibody level cannot be demonstrated in paired sera or when virus is not cultured.

Diagnosis of postnatally acquired rubella by use of three enzyme-linked immunosorbent assays for specific immunoglobulins G and M and single radial hemolysis for specific immunoglobulin G.

Three commercial indirect enzyme-linked immunosorbent assays (ELISAs) (RUBELISA, Enzygnost-Rubella, and Rubazyme) and a commercial single radial hemolysis (SRH) test (Rubazone) were evaluated for the diagnosis of acute rubella by testing 41 acute- (less than 7 days postonset) and convalescent-phase (8 to 82 days postonset) serum pairs from cases of rubella previously confirmed by significant change in the hemagglutination inhibition test titer. Specificity was tested by using 10 acute- and convalescent-phase serum samples from patients with rash not confirmed as rubella (control group). In testing for rubella-specific immunoglobulin G (IgG) antibody, Enzygnost-Rubella and Rubazyme confirmed infection in 40 and RUBELISA in 39 of the 41 proven rubella patients. For one patient all three ELISAs failed to show a significant titer rise. No false-positive diagnoses occurred in the control group, although a suspected infection was shown by Rubazyme in one patient. No specific IgM could be detected in this case. Single radial hemolysis confirmed infection in 39 of the 41 proven rubella patients, and one false-positive diagnosis occurred in the control group patients. Of the 43 convalescent-phase serum samples, rubella-specific IgM was detected in 42 by Enzygnost-Rubella, in 41 by RUBELISA M, and in 39 by Rubazyme-M. For a rapid diagnosis with acute-phase sera, specific IgM detection by ELISA was most reliable in hemagglutination inhibition test-positive sera; of 18 such serum samples IgM was shown in 15 by Enzygnost-Rubella, in 13 by RUBELISA M, and in 11 by Rubazyme-M. False-positive specific IgM results were shown by Rubazyme-M in two serum samples from one patient in the control group. These serum samples were negative with the other two ELISA methods.

Detection and persistence of specific IgM antibody to Coxiella burnetii by enzyme-linked immunosorbent assay: a comparison with immunofluorescence and complement fixation tests.

An enzyme-linked immunosorbent assay (ELISA) for specific IgM antibody to Coxiella burnetii was compared with the indirect immunofluorescence (FA) test in studies of 130 serum samples from 38 patients with Q fever. The IgM fractions obtained after rate zonal-density gradient ultracentrifugation of 37 serum samples from 12 patients were also studied in a complement fixation test (CF-DG). Specific IgM antibody to C burnetii was detected by all three methods in sera collected two to eight weeks after the onset of symptoms. The longest period for which specific IgM was shown to persist was 17 weeks by ELISA and FA and 10 weeks by CF-DG. The ELISA is performed with a single dilution of convalescent-phase serum and offers advantages over the subjective FA and the technically tedious CF-DG methods. The estimation of C burnetii-specific IgM by ELISA or FA is useful for the confirmation of infection when a rising titer of complement-fixing antibody cannot be demonstrated because acute-phase blood samples are not available.

Immunoglobulin responses to Coxiella burnetii (Q fever): single-serum diagnosis of acute infection, using an immunofluorescence technique.

Blood specimens were collected over various periods of time from 30 abattoir workers with a clinical diagnosis of Q fever. All specimens were tested for complement-fixing antibodies and for specific immunoglobulin M (IgM) globulins to phase 1 and 2 Coxiella burnetii organisms by an immunofluorescence technique. All 22 patients with increasing levels of complement-fixing antibodies were shown to have generated specific IgM globulins, as did 4 patients with high convalescent titers but from whom "acute" specimens were not collected. Four individuals who did not show increasing levels of complement-fixing antibodies did not produce measurable levels of specific IgM. All patients with Q fever gave positive specific IgM results by 2 weeks after the onset of symptoms. IgM to phase 1 antigen persisted for 27 weeks in one patient, but IgM to phase 2 antigen was not detectable beyond 17 weeks. The estimation of Q fever-specific IgM has proved useful in confirming infection when only a "convalescent" blood specimen is available.