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Application of O-serotyping in a study of Providencia rettgeri (Proteus rettgeri) isolated from human and nonhuman sources.

A somatic (O) antigen serotyping scheme for Providencia rettgeri (Proteus rettgeri) was modified to exclude O-type strains recently reclassified as urea-positive Providencia stuartii and was extended to include new serotypes to provide for serotyping on the basis of 93 O-antigens. Isolates from two hospitals, five public health laboratories, and nonhuman sources (polluted water and frogs) were serotyped. The 112 isolates collected from a large general hospital over a 99-month period were distributed among 42 O-serotypes. No serotype showed significant predominance that would suggest the occurrence of human strains that are more prone than others to cause human infections, but in an institution experiencing cross-infection, 11 of the 22 (50%) isolates belonged to one serotype. The 54 isolates from the five public health laboratories belonged to 33 serotypes, 15 of which were found also among hospital isolates. All but 5 of 99 frog isolates were typable, and the 94 typable isolates were separated into 25 serotypes. Each of the four isolates from polluted water samples was of a different serotype. Sixteen of the serotypes found in frogs and three found in water were also identified among human isolates.

Animals

Alternative quadruplex real-time PCR reactions for detection and discrimination of Streptococcus pneumoniae serotypes within serogroup 6.

UNLABELLED: Streptococcus pneumoniae causes significant morbidity and mortality worldwide, and serotyping is important to assess the burden of disease that is vaccine preventable. For serotyping, the Centers for Disease Control and Prevention (CDC) use a series of 12 real-time multiplex PCRs (rmPCRs) performed in quadruplex reactions; however, rmPCR reaction 5 (rmPCR-5) for serotypes 6A, 6B, 6C, and 6D often failed at low DNA concentrations. This study investigated the cause of rmPCR-5 failure and provided alternative rmPCRs to resolve this issue. Quadruplex rmPCR target sequences were compared to S. pneumoniae reference genomes. Reactions rmPCR-5 [6ABCD, 6AB, 6BD, and 6CD] and rm-PCR-11 [37, 10F, 11BC, and 18CFBA] were compared to alternative reactions rmPCR-A1 [6ABCD, 10F, 11BC, and 18CFBA] and rmPCR-A2 [37, 6AB, 6BD, and 6CD]. All rmPCRs were tested using 10-fold serial dilutions of DNA from representative serotypes, and analytical specificity was assessed using DNA from other S. pneumoniae serotypes or various streptococci and Gram-positive cocci. Failure of rmPCR-5 was associated with overlapping 6ABCD and 6BD targets. Separation of these targets in the alternative rmPCRs-A1 and rmPCR-A2 allowed sensitive and specific detection and discrimination of serotypes 6A, 6B, 6C, and 6D, without impacting the detection of serotypes 10F, 11BC, 18CFBA, and 37. This study highlights the importance of rigorous author and peer-review to avoid manuscript errors and unintended consequences. By explaining what caused rmPCR-5 failure and proposing alternative reactions rmPCRs-A1 and rmPCR-A2, this study demonstrates the value of scientific collaboration to ensure molecular assays best serve the scientific community. IMPORTANCE: Streptococcus pneumoniae is a bacterium that can cause life-threatening infections like pneumonia and meningitis, leading to millions of deaths worldwide each year. A key feature enabling S. pneumoniae to cause disease is its sugar coating, allowing it to avoid the immune system. These surface sugars are the target of S. pneumoniae vaccines. However, vaccines only protect against some sugars and understanding which ones are on the surface of S. pneumoniae is called "serotyping." The Centers for Disease Control and Prevention (CDC) have protocols that allow us to predict S. pneumoniae serotypes by looking at its DNA. We found errors in the CDC protocols and provided a simple solution to fix them. Ultimately, having accurate serotyping protocols allows us to know how much disease is preventable by vaccine, allows us to monitor how well vaccine are working, and helps develop new vaccines if needed.

Streptococcus pneumoniae

Role of protein serotype antigens in protection against disease due to Neisseria meningitidis.

Evidence is provided for the protective effects of antibody to noncapsular antigens in immunity to meningococcal disease. Serotype 2 strains were associated with 50% of disease due to group B Neisseria meningitidis in Europe. The amount of serotype antibody in normal individuals is related to age, and adult levels are reached by about 10 years of age. Children under two years of age who have recovered from disease due to groups B and CN. meningitidis of serotype 2 make antibody to serotype 2; this fact indicates the potential immunogenicity of the serotype antigen in young children. The isolated serotype 2 antigen injected into rabbits elicited bactericidal antibody that was only weakly opsonic. Guinea pigs that were infected via spring implants with serotype 2 N. meningitidis developed antibody to serotype 2. After clearance of infection these animals became resistant to challenge with serotype 2 strains of groups B and C N. meningitidis. This serotypes-specific immunity to reinfection persisted for at least four months.

Adult

Trends of serotypes and resistance among Streptococcus pneumoniae in the UK and Ireland (1999-2019).

OBJECTIVES: This study aimed to report the serotype distribution of Streptococcus pneumoniae isolates from UK and Irish patients with bacteraemia or community-associated lower respiratory tract infections (CA-LRTI). Depending upon the year, these were from 23 to 39 sentinel laboratories and were collected between 1999 and 2019, thus spanning the introduction of pneumococcal conjugate vaccines, PCV7 and PCV13. METHODS: Pneumococcal identification, susceptibility testing and serotyping were undertaken by a central laboratory. Changes in serotype distributions and among the predominant types showing antibiotic non-susceptibility were reviewed in relation to vaccine deployment. RESULTS: Following the introduction of PCV7 (2006) and PCV13 (2010), major shifts occurred in serotype prevalence for both bacteraemia and CA-LRTI. PCV7 types and most PCV13 types (but not 3 and 19A) were largely or wholly displaced. Many of the displaced types (e.g. 6B, 9V, 14, 19F and 23B) had been internationally prevalent and were associated with antibiotic resistance. Other serotypes-many included within the older pneumococcal polysaccharide vaccine, PPV23-expanded into the space, with serotype 8 becoming especially prominent in bacteraemia, though not respiratory infections. Further increasingly prevalent types included 9N, 10A, 12F and 22F. Serotype 15A, often multi-resistant, rose then fell in relative importance after deployment of PCV13. Among the currently most prevalent types, serotype 3 is rarely resistant to agents besides tetracyclines and bloodstream serotype 8 isolates mostly are fully susceptible. CONCLUSIONS: These data offer a comparison of serotypes associated with bacteraemia and respiratory disease over two decades in the UK and Ireland.

Humans

Epidemiologic differences among serotypes of Cryptococcus neoformans.

In the USA, the most prevalent serotype of the fungus, Cryptococcus neoformans, was serotype A. The serotype constituted 203 of 272 isolates from infections and 85 of 89 isolates from the environment. Serotype B or C isolates were infrequent causes of infection, except in Southern California, and were infrequent causes of infecand were not isolated at all from environmental sources. In Southern California, the absence of serotypes B and C in 67 soil and pigeon dropping isolates was striking, considering that 25 of 49 isolates from infections were serotypes B or C. The site in nature where serotypes B and C exist is currently unknown but differs from that of serotypes A and D. Serotype D may be unusually prevalent in both environmental and patient isolates from Denmark and Italy. Of 24 isolates from those countries, 21 were serotype D.

Animals

Yersinia enterocolitica: serotypes and biotypes isolated from humans and the environment in Quebec, Canada.

Thirty-one strains of Yersinia enterocolitica were isolated from food and surface water. During the period of January 1975 to June 1977, 157 strains from 143 human cases were also isolated. Among the different serotypes from nonhuman sources, serotypes 6,30 and 4,32 were the most common. Serotype 4,32 was present only in food and not in water. Serotype 3 was isolated only from humans. Among the different serotypes from human cases, 73.9% belonged to serotype 3. Only serotype 3 was isolated from children under 4 years old. The presence of other serotypes increased and that of serotype 3 decreased in frequency as the age progressed. No serotype 3 was isolated from human cases aged 50 years and more.

Adolescent

Haemophilus parahaemolyticus serotypes. Pathogenicity and cross immunity.

Pigs inoculated intranasally with Haemophilus parahaemolyticus, serotype 2, resisted challenge 3 weeks later with serotypes 2, 4 and 5 without showing clinical symptoms. The pigs were sacrificed 2 days after challenge, and post mortem examination showed a chronic pleuropneumonia from which only serotype 2 was re-isolated. Pigs inoculated intranasally with H. parahaemolyticus, serotype 2, showed no clinical symptoms when challenged 3 weeks later with serotype 1. Post mortem examination revealed a chronic pleuropneumonia with areas of necrosis from which H. parahaemolyticus, serotype 2, was re-isolated, but also small areas of a more acute fibrinous pneumonia from which serotype 1 was re-isolated. The control pig inoculated with only serotype 1 showed a severe acute fibrinous pleuropneumonia. The results indicate that a considerable cross immunity exists between the various serotypes of H. parahaemolyticus.

Animals

Haemophilus parahaemolyticus serotypes. Serological response.

Serotypes 1,2,4 and 5 of Haemophilus parahaemolyticus were inoculated into, respectively, 4,4,2 and 5 pigs. Serum samples were tested for circulating antibodies by the modified complement fixation test (CF test). When individual serotypes were used as antigen, titers were found only to the serotype which had been used for inoculation. Using antigen in which the serotypes were pooled, antibodies were demonstrated in sera from all the pigs. The CF titers obtained with the pooled antigen were equivalent to those found with each serotype separately. When the CF test was used for serological examination of field sera there was full agreement between the results obtained with the pooled antigen and those obtained with serotype 2 antigen alone. No cross reactions were found with the pooled antigen in herds that were sero-positive to Haemophilus parasuis, strain 4800. The experiment has shown that there is no serological cross reaction between serotypes 1, 2, 4 and 5 when they are used as antigen in the CF test. Also, the results imply that with a pool of the different serotypes of Haemophilus paralyticus as antigen similar results may be obtained as with the single serotype 2 antigen.

Animals

Heterogenity of serotypes of Neisseria meningitidis that cause endemic disease.

Three collections of strains of Neisseria meningitidis that caused meningococcal disease during nonepidemic periods were serotyped to determine whether serotypes that cause endemic disease are more heterogeneous than those responsible for epidemic disease. Thirty-four strains isolated from pediatric patients in Houston, Texas, from February 1977 to March 1978 were of three separate serogroups and 11 serotypes; 27 contemporary (1977-1978) strains from predominantly military populations, obtained nationwide, were of six serogroups and six serotypes, while 11 strains isolated at military posts in the southwest United States from 1970 through 1976 were of four serogroups and five serotypes. Between 9% and 20% of the strains were nontypable, while type II strains which were responsible for the epidemics in the Northern and Western Hemispheres earlier in the 1970's, accounted for only 20%-44% of the strains. In contast to epidemics, which appear to be caused by a single serotype, endemic meningococcal disease appears to be caused by a broad, heterogeneous distribution of serotypes. Thus, development of a serotype-specific vaccine may have limited application to the prevention of endemic meningococcal disease.

Adult

The dynamics of streptococcal infections in a defined population of children: serotypes associated with skin and respiratory infections.

Serial observations including cultures of the upper respiratory tract and of infected skin lesions and streptococcal antibody determinations were made over a two-year period in a semi-closed population of children between 10 months and 15 years of age. There was a high prevalence of group A streptococci in nose and throat cultures and of skin lesions containing these organisms. Almost 90% of the study population developed streptococcal impetigo during the study period. A slightly higher proportion of males than females developed skin infection but there was no relationship to age. Impetigo was observed throughout the calendar year, exceeding 12% of child-visits in one winter month, but was generally more common in the summer and fall. Conversely, group A streptococci were more often isolated from nose and throat cultures in the winter months. The increase in recovery of streptococci from nose and throat cultures lagged behind the increase in streptococcal impetigo and continued into the winter months, when the prevalence of impetigo had declined. Calculation of ratios for individual streptococcal serotypes isolated from different body sites revealed a clear cut distinction between "respiratory" and "impetigo" serotypes, with respect to both prevalence and acquisition rates. Respiratory serotypes were more commonly isolated in the winter and impetigo serotypes in the summer and fall. Significant antibody responses to extracellular antigens of the streptococcus were documented for pharyngeal acquisitions of both impetigo and respiratory serotypes and for skin lesions associated with impetigo serotypes. Group A streptococcal serotypes may be divided into three categories on the basis of their human pathogenicity for body sites: some with the potential for respiratory infection, others with the potential for skin infection and a few unusual serotypes with the potential for infecting both sites.

Age Factors

Heterotypic exclusion between vesicular stomatitis viruses of the New Jersey and Indiana serotypes.

Co-infection of cells with vesicular stomatitis viruses of the Indiana and New Jersey serotypes results in interference. Using specifically-labelled immunofluorescent antibodies, it was demonstrated that within any one co-infected cell, one virus serotype replicated to the relative exclusion of the other serotype. This result was further substantiated by an examination of the virus serotypes released by infectious centres co-infected with both viruses. Dominance of one serotype over the other was shown to be a function of the relative multiplicity of the two viruses. Superinfection by the second serotype at a higher multiplicity resulted in dominance by the second virus during the early period (up to 1-5 h) post-infection. After this time, the minority virus was able to overcome this dominance. Dominance of the majority virus was also abolished by u.v; inactivation. Cell protein synthesis appeared to be less affected in cells infected with both serotypes than when infection was with a single serotype.

Fluorescent Antibody Technique

The impact of the COVID-19 pandemic on the incidence of invasive pneumococcal disease in the Czech Republic and whole genome sequencing analysis of Streptococcus pneumoniae serotypes 3 and 19A from 2018-2024.

AIM: To describe in detail changes in the incidence of invasive pneumococcal disease in the Czech Republic during and after the COVID-19 pandemic. Another objective is molecular analysis of S. pneumoniae isolates of serotypes 3 and 19A recovered in the Czech Republic between 2018 and 2024. MATERIAL AND METHODS: Data on the incidence of invasive pneumococcal disease and S. pneumoniae serotypes were obtained from the invasive pneumococcal disease surveillance program in the Czech Republic. S. pneumoniae isolates of serotypes 3 (63) and 19A (66) from 2018-2024 were subjected to whole genome sequencing (WGS) to characterize the GPSCs (Global Pneumococcal Sequence Clusters) and STs (sequence types) and place them in a global context. RESULTS: Results: During the COVID-19 pandemic, a significant decline was observed in the incidence of invasive pneumococcal disease in the Czech Republic. Following the pandemic, the incidence of invasive pneumococcal disease rose again to significantly higher levels than before the pandemic. Compared to the 2018–2019 period, the incidence of certain serotypes increased in 2023–2024, including vaccine serotypes 3, 4, 14, and 15B, while the incidence of serotypes 8, 12F, and 15A, among others, decreased. Whole genome sequencing analysis demonstrated the dominance of GPSC12 ST-180 among serotype 3 isolates throughout the study period. Among serotype 19A isolates, GPSC4 prevailed, particularly ST-416. CONCLUSIONS: The COVID-19 pandemic has demonstrated how rapidly the epidemiological situation of invasive pneumococcal disease can change and that continuous, systematic surveillance of invasive pneumococcal disease is necessary. The best prevention against invasive pneumococcal disease is vaccination, primarily with higher valency pneumococcal conjugate vaccines.

Czech Republic

Antigenic structure and relationship between serotypes 1 and 2 of Haemophilus paragallinarum.

Antigenic structure and relationship between serotypes 1 and 2 of Haemophilus paragallinarum were analyzed by the rapid plate agglutination and cross-absorption tests. Encapsulated strain forming iridescent colony type of both serotypes 1 and 2 had at least three antigens: heat-labile and trypsin-sensitive (L), heat-labile and trypsin-resistant (HL), and heat-stable and trypsin-resistant (HS). The L was a major antigen located in a surface and divided serologically into three parts: L1, L2, and L3. The L1 was specifici to serotype 1, the L2 was specific to serotype 2, and the L3 was a common antigen shared by serotypes 1 and 2. The HL and HS were common antigens between serotypes. By trypsinization or heating at 121 C, L antigen lost its agglutinability and agglutinin-producing ability. Nonencapsulated organisms forming noniridescent colony type lacked the L antigen. These results suggested that antigenic structure of H paragallinarum serotypes 1 was L1, L3, HL and HS, while serotype 2 was L2, L3, HL, and HS.

Agglutination Tests

Special O:K:H serotypes among enterotoxigenic E. coli strains from diarrhea in adults and children. Occurrence of the CF (colonization factor) antigen and of hemagglutinating abilities.

O:H serotypes previously found to be prevalent among a number of toxigenic strains from several geographic areas were examined for polysaccharide K antigen type. Members of each O:H serotype had the same type of K antigen and were found to be characterized by a certain fermentation pattern. Some O:H serotypes had no K antigen. The serofermentative types defined were: 06:K15:H16, 08:K40:H9, 015:H11, 025:K7:H42, 025:K98:H-, 078:H11, 078:H12, and 0149:H10. Some strains of the last-mentioned serotype, which were suspected of having caused a food-borne infection, had K88. This serotype belongs to the group of strains causing diarrhea in swine. The surface antigen (CF) described as a colonization factor [5] was demonstrated in 078:H-, 078:H11, and 078:H12 strains; but not in any strain of the other serotypes nor in any of 248 strains belonging to 078 but not isolated from cases of human diarrhea. Presence of the CF antigen was correlated with presence of a mannose-resistant ability to cause agglutination of human red cells. Behavior of the other serotypes as regards hemagglutinating abilities was examined and 025:K7:H42 strains were found to be very similar to the 078 strains in this respect.

ABO Blood-Group System

A study of 666 strains of Candida albicans: correlation between serotype and susceptibility to 5-fluorocytosine.

The minimal inhibitory concentrations (MICs) of 5-fluorocytosine (5-FC) and the serotypes were determined for 666 strains of Candida albicans isolated from white patients in Canada. Of these strains, 495 (74.3%) were serotype A and 71 (25.7%) were serotype B. Resistance to 5-FC (MIC, greater than or equal to 25 microgram/ml) was observed in 11.3% of the serotype A strains and in 49.7% of the strains of serotype B. Most specimens were from the urogenital and respiratory tracts. The results showed the difference between the distribution of the two serotypes of C. albicans in North America and that in Europe. Serotyping of C. albicans strains by indirect immunofluorescence is a rapid means of predicting the de novo sensitivity of this year to 5-FC. The specificity of this technique for the isolated strains was approximately 79%.

Animals

Compatible results obtained from biotyping and serotyping in Serratia marcescens.

The correspondence between complete serotype and biotype (P.A.D. Grimont and F. Grimont, J. Clin. Microbiol. 8:73-83, 1978) of 474 Serratia marcescens strains was studied. Of 127 serotypes, 70 were represented by two or more strains of the same serotype belonged to one biotype. However, for 91% of serotypes, strains of the same serotype belonged to one biogroup--i.e., a group of closely related biotypes. Biogroups are A1 (A1a, A1b); A2/6 (A2a, A2b, A6a, A6b); A3 (A3a, A3b, A3c, A3d); A4 (A4a, A4b); A5/8 (A5, A8a, A8b, A8c); and TCT (TCT, TT). Only two serotypes were composed of a mixture of pigmented and nonpigmented biogroups. Pigmented biogroups (A1 and A2/6) were otherwise differentiated from nonpigmented biogroups (A3, A4, A5/8, and TCT) by serotyping. Some biogroups preferentially occurred in some O serogroups: A4 in 01; A2/6 in O6, O8, and O14; and A3 in O9, O12, and O15. Three H serogroups were found to be biochemically homogeneous: H1, H7, and H20 were respectively and uniquely composed of biogroups A4, TCT, and A3. A square matrix of O versus H serogroups, with the corresponding biogroup for each O X H combination, was used for comparisons between O groups and between H groups. Identical patterns of biogroups were shown by serogroups O6, O8, and O14. Taxonomical, ecological, and practical consequences of these findings are discussed.

Carbon

Demonstration of dual rhinovirus infection in humans by isolation of different serotypes in human heteroploid (HeLa) and human diploid fibroblast cell cultures.

The ability to isolate rhinoviruses in human heteroploid cell cultures was investigated by inoculating HeLa cells (HeLa M) with specimens previously shown to be positive in human diploid cell cultures. The 135 positive specimens selected were representative of 22 different rhinovirus types, and 4 to 9 specimens were available for each serotype. Specimens were inoculated into human diploid fetal tonsil fibroblasts (FT), HeLa cells with 30 mM Mg2+, and HeLa cells without increased Mg2+. One hundred twelve rhinovirus strains (83%) were reisolated in FT cells, whereas 76 rhinovirus strains (56%) were recovered in HeLa cells with 30 mM Mg2+. All strains recovered in FT were the same serotype as that originally recovered in diploid cells, but five of the HeLa cell isolates (3.7% of total specimens) were different serotypes, indicating dual rhinovirus infections. Four rhinovirus serotypes, (3, 42, 48, and 70) were recovered in HeLa but not in diploid cells; these serotypes were rare in our previous studies. Isolation of rhinovirus in FT cells was usually accomplished at first passage, whereas rhinovirus cytopathic effects in HeLa cells were not observed at first passage, but required one, two, or (rarely) three blind passages. Only 28 rhinoviruses (21%) were recovered in HeLa cells without increased Mg2+; however, three serotypes, types 16, 36, and 58, were recovered as effectively in HeLa cells, with or without added Mg2+, as they were in FT cells. In general, rhinoviruses were less efficiently recovered in HeLa cells; however, certain serotypes may be detected better by HeLa cells.

Cell Line

In vitro RNA transcription by the New Jersey serotype of vesicular stomatitis virus. I. Characterization of the mRNA species.

The in vitro RNA synthesis by the virion-associated RNA polymerase of vesicular stomatitis virus (VSV), New Jersey serotype, was compared with that of the serologically distinct Indiana serotype of VSV. The New Jersey serotype of VSV synthesized five distinct mRNA species in vitro, three of which were smaller than the corresponding species synthesized by the Indiana serotype of VSV. These included the mRNA's coding for the G, M, and NS proteins. By hybridization experiments, virtually no sequence homology was detected between the mRNA's of the two serotypes. Despite this lack of overall homology, the 12 to 18S mRNA species of both serotype contained a common 5'-terminal hexanucleotide sequence, G(5')ppp(5')A-A-C-A-G. The signicance of this finding in light of specific interactions between the two serotypes of VSV in vivo is discussed.

Base Sequence