Simultaneous detection of 33 Streptococcus suis serotypes using the luminex xTAG® assay™

Bai, Xuemei; Liu, Zhijie; Ji, Shaobo; Gottschalk, Marcelo; Zheng, Han; Xu, Jianguo

doi:10.1016/j.mimet.2015.07.018

Cited by 20 publications

(29 citation statements)

References 22 publications

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“…Chromosomal DNA was prepared by using a method described previously (15). The species identity of the 486 isolates was determined to be S. suis by amplification of the 16S rRNA, recN, and thrA genes (13,18,19).…”

Section: Methodsmentioning

confidence: 99%

“…Capsular gene typing methods developed in our laboratory were used to assign cps loci of 486 isolates (13,16,17,20). Known subtypes of NCL1, NCL2, NCL3, NCL7, and NCL8 were determined based on the variable presence of 13 genes and 4 transposase genes found in our previous study by PCR amplification (16) (Tables 1 and 2).…”

Section: Methodsmentioning

confidence: 99%

“…The 18-plex detection system was set up based on a protocol described in a previous study (13), and the primers used to amplify S. suis NCL-specific wzy target sequences are listed in Table 3. In brief, the unique "TAG" sequence and biotin label were specifically incorporated into wzy-specific amplification products in an mPCR reaction using cycling parameters of 94°C for 5 min and 30 cycles of 94°C for 30 s, 58°C for 30 s, and 72°C for 30 s, followed by a final elongation step at 72°C for 10 min.…”

Section: Methodsmentioning

confidence: 99%

“…The threshold of the detection limit was determined by using serially diluted DNA from a representative isolate of each NCL type. Non-S. suis isolates used in our previous study (13) were also used to determine the specificity of the system in the study. Two independent experiments were performed to establish the sensitivity and specificity of the system.…”

Section: Methodsmentioning

confidence: 99%

“…CPSs of all serotypes are thought to be synthesized by the Wzx/Wzy pathway. Based on serotype-specific wzy genes, 3 multiplex capsular gene typing systems have been developed to identify S. suis serotypes (13)(14)(15). These systems allow the simultaneous detection of multiple nucleic acid sequences in a single reaction and can greatly reduce the time, cost, and work associated with conventional serotyping technologies.…”

mentioning

confidence: 99%

See 4 more Smart Citations

Novel Capsular Polysaccharide Loci and New Diagnostic Tools for High-Throughput Capsular Gene Typing in Streptococcus suis

Qiu

Bai

Lan

et al. 2016

Appl Environ Microbiol

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Streptococcus suis is an important pathogen of pigs and may cause serious disease in humans. Serotyping is an important tool for detection and epidemiological studies of S. suis. Thirty-three reference serotypes and nine novel cps loci (NCLs) are recognized in S. suis. To gain a better understanding of the prevalence and genetic characteristics of NCLs, we investigated the serotype identity of 486 isolates isolated between 2013 and 2015 in China by capsular gene typing methods. Two hundred seventy-six isolates carried NCLs belonging to 16 groups, 8 of which appear to have not been reported previously. These isolates showed autoagglutination, polyagglutination, or nonagglutination with reference antisera and thus were nonserotypeable. Almost all isolates carrying the unknown NCLs were encapsulated, with various capsular thicknesses, indicating that they are most likely novel serotypes. To simultaneously identify the currently recognized 17 NCLs, an 18-plex detection system using the Luminex xTAG universal array technology was developed. Our data also provide valuable genetic information for monitoring the variations within NCLs by investigating the genetic characteristics of different subtypes within NCLs. IMPORTANCENonserotypeable Streptococcus suis isolates have been reported in many studies, and 9 novel cps loci (NCLs) have already been identified in nonserotypeable isolates. Moreover, novel cps loci are continually being found. The main purpose of this study was to investigate the prevalence and characteristics of NCLs in S. suis isolates recovered between 2013 and 2015 in China. This study provides valuable genetic information for monitoring the variations within NCLs. Meanwhile, a fast and cost-effective 18-plex detection system that can simultaneously identify the currently recognized 17 NCLs was developed in this study. This system will serve as a valuable tool for detecting known and identifying additional novel cps loci among nonserotypeable S. suis isolates. Streptococcus suis is an important pathogen of pigs and may cause serious disease in humans (1-3). Clinically healthy pigs can carry S. suis in their nasopharynx, contributing to the dissemination of this pathogen (4, 5). Moreover, a potential role of isolates from healthy pigs is as donors to transfer the new adaptive phenotypes to pathogenic isolates (6). The capsular polysaccharide (CPS) shields S. suis from host phagocytes and is a major virulence factor (7). Antisera to CPSs are used to distinguish antigenic differences among them. Serotyping is also an important epidemiological method for S. suis surveillance. This reflects the importance of including isolates from healthy pigs to better understand the diversity and evolution of CPSs in S. suis populations.A total of 35 serotypes (types 1 through 34 and type 1/2) of S. suis have been identified by different studies in the 1980s and 1990s (8-11). In 2005, strains of serotypes 32 and 34 were reclassified as Streptococcus orisratti (12). The CPS synthesis genes are known to be clustered ...

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

Novel Capsular Polysaccharide Loci and New Diagnostic Tools for High-Throughput Capsular Gene Typing in Streptococcus suis

Qiu

Bai

Lan

et al. 2016

Appl Environ Microbiol

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Streptococcosis

Gottschalk¹,

Segura²

2019

Diseases of Swine

116

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Prototype of Multiplex Bead Assay for Quantification of Three Serum Biomarkers for In Vitro Diagnosis of Endometriosis

Signorile¹,

Baldi²

2016

Journal Cellular Physiology

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Endometriosis is a very common disease, affecting 10% of women in the reproductive age. To date, a significant delay between onset of the symptoms and definitive diagnosis is caused by the lack of a reliable non-invasive diagnostic test. Recently, the potential value as diagnostic markers for endometriosis of three proteins (Zn-alpha2-glycoprotein, serum albumin, and complement C3 precursor), has been showed. In this article, we have defined the experimental conditions for the development of a multiplex bead array assay for rapid and simultaneous quantification of these three biomarkers in the serum of patients with endometriosis. Finally, pivotal experiments on a small cohort of patients have confirmed the diagnostic value of this assay. J. Cell. Physiol. 231: 2622-2627, 2016. © 2016 Wiley Periodicals, Inc.

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Simultaneous detection of 33 Streptococcus suis serotypes using the luminex xTAG® assay™

Cited by 20 publications

References 22 publications

Novel Capsular Polysaccharide Loci and New Diagnostic Tools for High-Throughput Capsular Gene Typing in Streptococcus suis

Novel Capsular Polysaccharide Loci and New Diagnostic Tools for High-Throughput Capsular Gene Typing in Streptococcus suis

Streptococcosis

Prototype of Multiplex Bead Assay for Quantification of Three Serum Biomarkers for In Vitro Diagnosis of Endometriosis

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