Evaluation of serological tests for diagnosis of Bordetella pertussis infection in adolescents and adults

Background and objective: Bacterial agglutination antibodies against Bordetella pertussis, Yamaguchi and Tohama strains, are frequently measured for serodiagnosis of pertussis infection in Japan. To determine the serological criteria, the comparative titres of bacterial agglutination antibody and anti‐pertussis toxin (PT) antibody were evaluated. Methods: Antibody titres were analysed in 36 definitive (fourfold increase in agglutination antibody) and 137 presumptive (high titre of single‐antibody) cases of B. pertussis infection among adolescents and adults, and in a control group of 318 healthy volunteers. Results: When a single Yamaguchi agglutinin titre of ≥1:1280 (> three SD above the geometric mean for the control group) was taken as diagnostic, the sensitivity and specificity at 4–5 weeks after onset of cough were 58% and 98%, respectively. Using this criterion, the clinical findings in presumptive cases were almost identical to those in definitive cases. When the two tests were compared using 318 control sera, there was no association between the Tohama agglutinin titre and the anti‐PT antibody titre, whereas a weak association between the Yamaguchi agglutinin titre and the anti‐PT antibody titre was observed. When the numbers of pertussis cases with high antibody titres in the two tests were compared, 60% of cases with a Yamaguchi agglutinin titre of ≥1:1280 showed an anti‐PT antibody titre of ≥100 EU/mL. Conclusions: These results indicate that the bacterial agglutination test is a method with low sensitivity and specificity for the diagnosis of B. pertussis infection. Therefore, to yield an accurate diagnosis, anti‐PT antibody levels should be measured instead of bacterial agglutination antibody.     

Frequency of serological evidence of Bordetella infections and mixed infections with other respiratory pathogens in university students with cough illnesses.

Banked acute-phase and convalescent-phase serum samples from a previous study of respiratory illness in university students were examined for significant (>/=2-fold) increases in ELISA titers of IgA and IgG antibody to Bordetella pertussis filamentous hemagglutinin, pertactin, and fimbriae-2 and >/=4-fold titer increases to agglutinogens by agglutination. ELISA titers of antibody to pertussis toxin could not be determined because of technical problems. Chlamydia pneumoniae infections were diagnosed by culture or by a >/=4-fold increase in immunofluorescence assay titer or a single high titer (>/=512). Mycoplasma pneumoniae, influenza A and B, adenovirus, and respiratory syncytial virus infections were diagnosed by >/=4-fold increases in complement fixation titer or a single high titer (>/=64). There were 319 subjects with cough of >/=5 days' duration, and of these, 47 (15%) had significant increases in antibody to B. pertussis antigens; 26 (8%) had significant increases to fimbriae-2 or agglutinogens, indicative of B. pertussis infection, and 2 (1%) had evidence of non-B. pertussis bordetella infections. Seventeen (36%) had evidence of mixed infections or cross-reacting antibodies (influenza B infections, 5; adenovirus infections, 4; influenza A infections, 3; C. pneumoniae infections, 3; and M. pneumoniae infections, 2). Our findings suggest that bordetella infections are common in young adults with cough illnesses (incidence, 9%), and a surprising number of these are mixed infections with other respiratory pathogens.     

Serological diagnosis of pertussis: IgM, IgA and IgG antibodies against Bordetella pertussis measured by enzyme-linked immunosorbent assay (ELISA)

Igm, IgA and IgG antibodies against Bordetella pertussis were measured by enzyme-linked immunosorbent assay (ELISA) with an ultrasonicate of formalin-killed bacteria (a mixture of strains 1, 2 and 1, 2, 3) as antigen and disposable polystyrene 9-cuvette blocks as the solid phase. The specificity properties of the assay were assessed by an inhibition technique. Of the microbes tested, only B. parapertussis was able to cause a significant inhibition. In addition, IgM and IgA antibodies against B. pertussis were only found in some sporadic cases of respiratory infections caused by other microbes. Sera, nasal swabs and cough plates were received from 198 patients with suspected whooping-cough. ELISA determinations were mostly made from only one serum sample of each patient. Paired sera were studied only from the culture-positive infants under 3 months of age. The number of positive cultures was highest in group under 3 months of age (41%), where the frequency of positive ELISA was lowest (20%). The use of paired sera strikingly increased the number of ELISA-positive individuals in this youngest patient group. In later life, the relationship between these tests changed: isolation was positive in only about 10% of the patients, whereas 29-64% yielded positive titres in ELISA. This study shows that pertussis ELISA is a valuable aid in the rapid diagnosis of pertussis, particularly of the atypical forms of the disease which mostly are culture-negative.     

Frequency of pertussis in children with prolongued cough

To determine the frequency of pertussis in children < or = 16 y who had prolonged cough (> or = 14 d), a prospective study was conducted at an outpatient clinic of a paediatric hospital. Nasopharyngeal swabs were taken for culture and nucleic acid testing by polymerase chain reaction (PCR) for Bordetella pertussis. Immunoglobulin A and immunoglobulin G antibodies against pertussis toxin (PT) were tested by ELISA in paired serum samples. A total of 148 patients were recruited during 1 y. Pertussis was detected in 25 (16.9%) patients with at least 1 of the tests. PCR was positive in 12 patients, and 9 cases was diagnosed serologically. Both PCR and serology were positive in 4 children. Duration of cough was longer in the patients with pertussis (median 33 vs 20, p = 0.03). Seropositivity of pertussis toxin was higher in pertussis negative patients during enrollment (24% vs 65%, p = 0.005). From the results of this study, B. pertussis seems to be common in our population despite high immunization rates with whole cell vaccine. Although the duration of cough is defined as longer than 21 d in some studies for pertussis case definition criteria, it was shorter than this in 3 of our cases.     

Evaluation of serology and nasopharyngeal cultures for diagnosis of pertussis in a vaccine efficacy trial

Nasopharyngeal cultures and titer rises in paired sera were evaluated in a placebo-controlled pertussis vaccine efficacy trial. IgG ELISA for filamentous hemagglutinin (FHA) identified 30 (88%) of 34 placebo recipients and 33 (89%) of 37 vaccine recipients with culture-verified Bordetella pertussis infections, whereas IgG ELISA for pertussis toxin (PT) showed higher diagnostic sensitivity in the placebo group than in the vaccine groups. The CHO cell assay did not improve sensitivity. Children with Bordetella parapertussis infections had rises of titers of antibody to FHA of the same magnitude as children with B. pertussis infections. Sensitive serologic criteria, based on the intraassay variations, identified 105 additional culture-negative cases with significant titer rises in paired sera. IgG ELISA for FHA and PT and IgA ELISA for FHA were reliable assays, and bacterial isolation rates were lower in vaccine recipients than in placebo recipients with serologically defined pertussis.     

Serologic diagnosis of pertussis: evaluation of pertussis toxin and other antigens in enzyme-linked immunosorbent assay

IgM, IgA, and IgG antibodies to Bordetella pertussis were measured in paired sera from 34 patients who were culture-positive for pertussis by enzyme-linked immunosorbent assay (ELISA) with disrupted B. pertussis bacteria, purified pertussis toxin, or outer membrane proteins (OMP) as antigens. Paired sera from 50 patients with other respiratory infections were used as controls. The sensitivities of the assays from paired sera were 61%, 90%, and 90% and specificities were 98%, 92%, and 72%, respectively. Of the patients culture-positive for pertussis, 68% had positive levels of antibody to pertussis toxin antigen in their first serum samples, obtained at the same time as samples for culture. Infants had antibody responses to pertussis toxin antigen, in contrast to weak antibody responses measured by B. pertussis antigen. The results from this study indicate that ELISA, especially measuring pertussis toxin IgA, is a valuable additional tool for diagnosing pertussis and can be used as a complementary test with cultures.     

Morbidity of pertussis in adolescents and adults

The effect of age on the clinical presentation of pertussis was assessed in 664 adolescent and adult cases. Complications were more frequent in adults than in adolescents (28% vs. 16%). Pneumonia occurred in 2% of patients <30 years old but in 5%-9% of older patients. Urinary incontinence occurred in 34% of women >/=50 years old. Duration of cough, risk of sinusitis, and number of nights with disturbed sleep increased with smoking and asthma. The secondary attack rate in other household members >/=12 years was 11%. Pertussis in secondary case patients was less severe than in index case patients but presented with classic symptoms. The main source of infection in adolescents was schoolmates or friends; in adults it was workplace or their children. Teachers and health care workers had a greater risk of pertussis than did the general population. The burden of disease appears to increase with age, with smoking, and with asthma.     

Serologic diagnosis of pertussis: comparison of enzyme-linked immunosorbent assay and bacterial agglutination

An enzyme-linked immunosorbent assay (ELISA) and bacterial agglutination (BA) method for determining the presence of antibodies to Bordetella pertussis were compared on serum samples from 21 patients with whooping cough and their 76 family members. The overall diagnostic agreement between the two methods was 77%. The data for BA-detected antibodies correlated best with IgG and IgA antibodies to B. pertussis. All of the culture-positive patients showed serologic positivity in both assays during the follow-up. Pertussis was diagnosed by ELISA in most cases from the first serum sample. Both methods proved to be good diagnostic aids in culture-negative patients, although the value of BA is more retrospective because of the need for paired sera. The kinetics of IgM, IgA, IgG, and agglutinating antibodies to B. pertussis is presented.     

Evidence of efficacy of the Lederle/Takeda acellular pertussis component diphtheria and tetanus toxoids and pertussis vaccine but not the Lederle whole-cell component diphtheria and tetanus toxoids and pertussis vaccine against Bordetella parapertussis infection.

A subanalysis of a recent cohort efficacy trial of a pertussis vaccine was performed to determine its efficacy against cough illnesses due to Bordetella parapertussis infections. Infants received four doses of either the Lederle/Takeda acellular pertussis component diphtheria and tetanus toxoids and pertussis (DTaP) vaccine or the Lederle whole-cell component diphtheria and tetanus toxoids and pertussis (DTP) vaccine at 3, 4.5, 6, and 15-18 months of age; controls received three doses of diphtheria and tetanus toxoids (DT) vaccine only. All subjects were prospectively followed for cough illnesses of > or = 7 days' duration; cases of B. parapertussis infection were confirmed by positive culture, household contact, or serology. Seventy-six cough illnesses due to B. parapertussis were identified; 24 occurred in 929 DTaP recipients, 37 in 937 DTP recipients, and 15 in 321 DT recipients, resulting in an efficacy of 50% for DTaP vaccine (95% CI [confidence interval], 5% to 74%) and 21% for DTP vaccine (95% CI, -45% to 56%). The data in the present analysis suggest that the Lederle/Takeda DTaP vaccine but not the Lederle whole-cell component DTP vaccine has efficacy against B. parapertussis infection.     

Diversity of etiological agent associated with aseptic meningitis--a survey on an epidemic in Tajimi City, Gifu Prefecture in 1984

During the period from May to September in 1984, an epidemic of aseptic meningitis occurred in Tajimi city, Gifu prefecture. Virological and serological investigations were performed. Cases involved ranged from 0 to 12 years of age, and 83.1% of them were under 7 years of age. Four types of enteroviruses were isolated from 38 to 59 cases (64.4%), and virus type isolated were coxsackie B1 (15 cases), B4 (2 cases), B5 (4 cases), echovirus 16 (14 cases), a mixture of coxsackie-virus B4 and echovirus 16 (2 cases) and an unidentified strain (one cases). Virus types isolated from cerebrospinal fluid were coxsackievirus B1 and B5. In the serological studies of 47 paired sera of patients, a significant rise in titer against 4 virus types isolated was 29.8% with coxsackievirus B1, 12.8% with coxasackievirus B4, 17.0% with coxsackievirus B5, and 34.0% with echovirus 16. In relation between virus type isolated and a significant rise in titer of neutralizing antibody, almost all of the patients' sera (80.0%) exhibited a significant rise in titer of neutralizing antibody against one isolated virus type, but some patient's sera (20.0%) exhibited a significant rise against two or more of the viruses.     

支气管哮喘患者记忆性T淋巴细胞对B淋巴细胞产生IgE作用的研究

目的 探讨记忆性（ＣＤ４＾＋ＣＤ４５ＲＯ＾＋）Ｔ淋巴细胞在支气管哮喘的发病中的作用。方法 分别分离出支气管哮喘病人及健康对照的ＣＤ４＾＋ＣＤ４５ＲＯ＾＋Ｔ淋巴细胞亚群,并将其与各自的Ｂ淋巴细胞共同培养,设定刺激组（美洲商陆有丝分裂原）及非刺激组,测定培养上清液中ＩｇＥ的含量。结果 哮喘病人自然状态的ＣＤ４＾＋ＣＤ４５ＲＯ＾＋Ｔ淋巴细胞对Ｂ淋巴细胞产生ＩｇＥ有正向促进作用,但有接受美洲商陆有丝分裂原     

Case report of collagen lung in SLE presenting with cough variant asthma: relation between the localization of responsible receptors and cough]

A 27-year-old nonsmoking woman complained of cough and chest oppression for two years since an episode of pneumonia. Clinical tests showed decrease in FEV1.0 during attacks of coughing and evidence of bronchial hypersensitivity. While these events fitted the picture of bronchial asthma, the nonwheezing cough suggested cough variant asthma. Antinuclear antibody and anti-ds DNA antibody were increased and leukopenia was recognized, suggesting the diagnosis of systemic lupus erythematosus (SLE). Bronchoalveolar lavage showed lymphocytic alveolitis and decreased T4/T8. These results were suggestive of collagen lung induced by SLE. Inhalation challenge with capsaicin and rapid intravenous injection of lobelin and alinamin indicated that peripheral c-fiber receptors were involved in the induction of coughing. We conclude that the peripheral lesion of collagen lung stimulates the peripheral c-fiber receptors, leading to cough variant asthma.     

Pertussis is a frequent cause of prolonged cough illness in adults and adolescents.

Although pertussis is increasingly recognized as a cause of prolonged cough illness in adolescents and adults, its prevalence is not well established. We evaluated pertussis infection in 442 adolescents and adults > or = 12 years old (mean age, 41.3 years) who had a cough-related illness of 7--56 days' duration. For 4 patients (0.9%), results of nasopharyngeal culture or PCR were positive for Bordetella pertussis; for 10 patients (2.3%), either results of culture or PCR were positive or pertussis antibody titers increased 4-fold. Eighty-eight patients (19.9%) had either laboratory-confirmed pertussis or laboratory evidence of pertussis. These patients had significantly longer duration of cough than did patients without laboratory evidence of pertussis (56 days vs. 46 days), and more of them had vomiting with cough (45.5% vs. 28.5%, respectively). Pertussis is a common cause of prolonged cough illness in adolescents and adults and is frequently associated with other symptoms of whooping cough.     

Bordetella pertussis, Bordetella parapertussis, Mycoplasma pneumoniae, Chlamydia pneumoniae and persistent cough in children.

Material collected during a prospective pertussis vaccine trial in 1992-95 was examined for Bordetella pertussis (culture and serology), Bordetella parapertussis (culture), Mycoplasma pneumoniae and Chlamydia pneumoniae (PCR). From 64% (99/155) of episodes with cough for less than 100 d, 115 aetiological agents were identified in one southern and one northern subset of DT-recipients. The most common single agent was B. pertussis, representing 56%(64/115), with a median cough period of 51 d, followed by M. pneumoniae 26%(30/115), 23 d, C. pneumoniae 17% (19/115), 26 d, and B. parapertussis 2% (2/115). For co-infections, the median duration of cough was about 60 d. Spasmodic cough for 21 d or more (clinical WHO criteria for pertussis) was present in 82% (41/50) of infections with B. pertussis as single agent, 38% (17/45) with B. parapertussis, 38% (5/13) with C. pneumoniae, 26% (5/19) with M. pneumoniae and 30%(17/56) in cases where no aetiology was found. In children with cough for more than 100 d (n = 78) using all vaccine arms, B. pertussis was responsible in 83% (65/78), in 21%(16/78) together with other agents. Acellular vaccines were more efficient against serious disease than whole cell vaccine. Antibiotic treatment was more common at the southern (34%) study site than at the northern one (12%). The findings indicate that diagnosis should rely on laboratory confirmation, both for rational treatment of an individual case and for monitoring outbreaks.     

呼出气一氧化氮检测在慢性咳嗽病因诊断中的价值

目的评估呼出气一氧化氮（FeNO）检测在慢性咳嗽患者病因诊断中的价值。方法选取2009年1月～2009年4月因慢性咳嗽在我院进行支气管激发试验的患者43例,先后予以FeNO、支气管激发试验检查。结果18例患者支气管激发试验阳性,诊断为咳嗽变异性哮喘。25例激发试验阴性患者,为非哮喘的慢性咳嗽。哮喘组患者FeNO水平显著高于非哮喘组,哮喘组患者FeNO值与LogPD20FEV1存在显著负相关性（r=-0．884）。FeNO诊断咳嗽变异性哮的最佳阈值为15．85ppb,其约登指数为0．753,灵敏度为83．3％,特异度为92．0％,阳性预计值为88．2％,阴性预计值88．5％。结论FeNO对咳嗽变异性哮的诊断具有较高的灵敏度和特异度,在慢性咳嗽的病因诊断中具有重要的应用价值。     

Human parainfluenza virus type 3 (HPIV 3) community-acquired pneumonia (CAP) mimicking pertussis in an adult: the diagnostic importance of hoarseness and monocytosis

Background

There are relatively few causes of acute community-acquired pneumonias (CAPs) in adults associated with prolonged cough. In adults the most common acute CAPs with a prominent and persistent nonproductive cough are due to Mycoplasma pneumoniae, Chlamydophilia (Chlamydia) pneumoniae, or Bordetella pertussis (pertussis). Pertussis is an underrecognized and underappreciated cause of CAP in adults. Different from classic pertussis in children, pertussis in adults presents with prolonged dry cough, that is, the “100-day cough.” In pertussis, the characteristic nonspecific laboratory findings are leukocytosis and relative lymphocytosis. Dry cough accompanied by hoarseness with CAP in an adult should suggest C. pneumoniae or a respiratory virus (eg, influenza, parainfluenza, respiratory syncytial virus).

Methods

We present the case of a young woman who presented with a prominent and persistent pertussis-like cough with hoarseness. She had no leukocytosis or relative lymphopenia, which argued against the diagnosis of pertussis. Notably, she had persistent monocytosis. Her protracted pertussis-like cough that persisted during her hospitalization was so impressive that the diagnostic impression was pertussis. Direct fluorescent antibody (FA) and throat cultures were negative for pertussis. Furthermore, her hoarseness suggested the possibility of C. pneumoniae, but her C. pneumoniae immunoglobulin-M titer was negative.

Results

Because C. pneumoniae was ruled out, her hoarseness suggested a respiratory viral cause. A respiratory FA viral panel and viral throat cultures were obtained. The respiratory FA viral panel was negative for influenza A/B, respiratory syncytial virus, metapneumovirus, adenovirus, cytomegalovirus, and parainfluenza viruses. However, her viral throat cultures grew parainfluenza virus type 3 (HPIV 3), confirming the diagnosis.

Conclusion

To the best of our knowledge, this is the first case of HPIV 3 CAP presenting with a prominent and persistent pertussoid cough in an adult mimicking pertussis with hoarseness and monocytosis.     

支气管哮喘、咳嗽变异性哮喘及急性支气管炎气道反应性特点及意义

目的探讨支气管哮喘、咳嗽变异性哮喘及急性支气管炎气道反应性特点,以便为临床诊断提供依据。方法采用日本产ＡＳＴＯＧＡＰＨＴＣＫ６０００ＣＶ气道反应测定仪,以乙酰甲胆碱为气道激发剂,观察６０例支气管哮喘、５８例咳嗽变异性哮喘及３７例急性支气管炎患者气道反应性变化。结果支气管哮喘和咳嗽变异性哮喘病人气道激发试验均为阳性,哮喘病人的气道反应阈值（Ｄｍｉｎ）低于咳嗽变异性哮喘病人（Ｐ＜００５）。急性支气管炎病人中,气道激发试验３３例阴性,占８９％,４例阳性,占１１％。４例阳性急性支气管炎患者的气道反应性曲线与哮喘组及咳嗽变异性哮喘组明显不同,其Ｄｍｉｎ也显著高于哮喘组（Ｐ＜００１）及咳嗽变异性哮喘组（Ｐ＜００５）。结论气道反应性测定对于不同类型哮喘及急性支气管炎的鉴别和指导治疗具有很好的临床应用价值。     

Immunogenicity of trivalent-inactivated influenza vaccine among children less than 4 years old

We studied the immunogenicity of trivalent-inactivated influenza vaccine. Subjects were 259 children under 4 years old who visited six pediatric clinics to undergo influenza vaccination. Age distribution was 64 aged <1.0, 65 aged 1.0-1.9, 64 aged 2.0-2.9, and 66 aged 3.0-3.9 years, including subjects who had been previously vaccinated within the last three years, 0% (0/64) aged <1.0, 26% (17/65) aged 1.0-1.9, 72% (46/64) aged 2.0-2.9, and 77% (51/66) aged 3.0-3.9 years old. Two doses of vaccine were given subcutaneously four weeks apart. Dosage was 0.l mL for children under 1 year old, while for children aged one year or older, dosage was 0.2mL, based on standard Japanese recommendations. To measure hemagglutination inhibition (HI) antibody titer, triplet sera were obtained before vaccination (S0), 4 weeks after the first vaccination (S1), and 4 weeks after the second vaccination (S2). The geometric mean of HI antibody titer, the response proportion (titer rise > or =4-fold), and the achievement proportion (postvaccination titer > or =1 : 40) were calculated by age group. Analysis of variance was used to estimate the independent effect of age and prevaccination titer on antibody increase. The geometric means of HI antibody titer were lower among the two younger age groups than among the two older age groups, regardless of vaccine strain or when blood samples were collected. The achievement proportion after 2 doses of vaccine in the <1.0, 1.0-1.9, 2.0-2.9, 3.0-3.9 year age groups were 38%, 58%, 89%, and 85% against A (HI) ; 52%, 54%, 81%, and 73% against A (H3) ; and 23%, 49%, 67%, and 71% against B. Regarding the analysis of variance, prevaccination titer consistently indicated strong effects on antibody increase, regardless of vaccine strain or combination of paired sera. After two doses of vaccine (S2/S0), significant effects of age on antibody induction were shown against A (H1) and B (p = 0.000 and 0.002). Thus, the immunogenicity of trivalent-inactivated influenza vaccine was strongly influenced by prevaccination titer and age. Even two doses of vaccine did not induce a protective antibody level in about 50 to 80% of subjects among infants aged <1.0 year, and 40 to 50% among children 1.0-1.9 year old.     

Whooping cough in Pakistan: Bordetella pertussis vs Bordetella parapertussis in 2005-2009

Pertussis, or whooping cough, is an acute respiratory disease mainly affecting infants and children and is caused by Bordetella pertussis and Bordetella parapertussis. The aim of this study was to investigate the share of Bordetella species from potential whooping cough cases during 2005-2009. Eight hundred and two samples from suspected pertussis cases were collected, mainly from 2 provinces of Pakistan. Bacterial culture, identification, DNA extraction and routinely used polymerase chain reaction (PCR) methods using IS1001, IS1002 and IS481 were used to identify the Bordetella species. The results were unexpected, because all of the isolates collected from the different cities were identified as B. parapertussis (7.4%); B. pertussis was not isolated from any sample. However, PCR results indicated the presence of a small percentage (0.6%) of B. pertussis among the total cases studied. This study suggests that vaccines to protect against both B. pertussis and B. parapertussis should be considered.     

Viral antibody studies in pancreatic disease.

Viral studies were performed on sera from 54 patients with recent acute pancreatitis, 10 with recurrent acute pancreatitis, seven with chronic pancreatitis, and 10 with pancreatic carcinoma, and on sera from 81 age- and sex-matched controls. In 29 of the acute pancreatitis patients from whom paired sera were obtained no convincing evidence of recent viral infection was found. A higher incidence of raised antibody titres against Coxsackie B3 and B4 was observed in the group of acute pancreatitis patients compared with their controls. The possible signficance of these observations and their relationship to the aetiology of the pancreatitis and to other immunological findings are discussed.