Variable number of tandem aminoacid repeats in adhesion-related CDS products in Mycoplasma hyopneumoniae strains

Castro, Luiza Amaral de; Pedroso, Thiago Rodrigues; Kuchiishi, Suzana Satomi; Ramenzoni, M. L. F.; Kich, Jalusa Deon; Zaha, Arnaldo; Vainstein, Marilene Henning; Ferreira, Henrique Bunselmeyer

doi:10.1016/j.vetmic.2006.04.022

Cited by 44 publications

(44 citation statements)

References 30 publications

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“…On the other hand, strains having 40 and 43 VNTR copy number in P97R1 were identified only in healthy lungs. The presence of nine repeats in M. hyopneumoniae strain J (CASTRO et al, 2006), known to be avirulent and have a low adherence activity, (VASCONCELOS et al, 2005), confirms the assumption that other determinants, like putative adhesins P76, P146 and P216, are involved in the adhesion process (CASTRO et al, 2006). Also, VNTR P97R2 is suggested as a required determinant for binding to the host respiratory surfaces, (JENKINS et al, 2006).…”

Section: Discussionsupporting

confidence: 52%

“…Differences in the number of aminoacidic repetitions due to the variability in the number of repetitive units contained in the VNTR could have implications in structural, physiochemical and antigenic characteristics of the corresponding proteins (CASTRO et al, 2006). However, there are no studies that show any relationship between such aminoacidic repetitions and the differences in pathogenicity of some M. hyopneumoniae strains.…”

Section: Discussionmentioning

confidence: 99%

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Detection, quantification and genetic variability of Mycoplasma hyopneumoniae from apparently healthy and pneumonic swine

Pulgarón¹,

Marques

Campos

et al. 2015

Braz. J. Vet. Res. Anim. Sci.

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Molecular differences among Mycoplasma hyopneumoniae strains present in pneumonic lungs of swine have been largely studied. However, no comparative studies concerning the strains present in apparently healthy pigs have been carried out. This study aimed to detect, quantify and perform molecular analysis of M. hyopneumoniae strains in pig lungs with and without pneumonic lesions. The detection of M. hyopneumoniae was performed using multiplex PCR (YAMAGUTI, 2008), real-time PCR (STRAIT et al., 2008) and multiple VNTR amplification (VRANCKX et al., 2011). Molecular characterization of the strains was achieved by analysis of the VNTR copy number in P97R1, P146R3, H2R1 and H4. M. hyopneumoniae was detected in samples from healthy and pneumonic pigs and the amount of M. hyopneumoniae positive samples detected varied with the type of assay. The greater number of positive samples was identified by the multiple VNTR amplification combined with capillary electrophoresis. Using real-time PCR, 4.9*10 4 M. hyopneumoniae genome copies/mL was detected in apparently healthy lungs. A mean quantity of 3.9*10 6 M. hyopneumoniae genome copies/mL was detected in pneumonic lungs. The analysis of VNTR copy number demonstrated a high genetic variability of the M. hyopneumoniae strains present in apparently healthy and pneumonic lungs. Strains having 3 VNTR copy number in P97R1, were detected only in pneumonic lungs and strains having 40 and 43 VNTR copy number in P146R3 were detected only in apparently healthy lungs. Despite the genetic variability of M. hyopneumoniae, predominant strains in the swine farms could be identified.Keywords: M. hyopneumoniae. Enzootic pneumonia. Healthy. Quantification. VNTR. ResumoAs diferenças moleculares entre as estirpes de Mycoplasma hyopneumoniae presentes em pulmões de suínos com pneumonia têm sido estudadas. Porém, estudos comparativos relativos às estirpes presentes nos suínos aparentemente saudáveis não foram levados a cabo. O objetivo do estudo foi a detecção, quantificação e análise molecular de M. hyopneumoniae nos pulmões suínos com e sem lesões pneumônicas. Para a detecção de M. hyopneumoniae usaramse o PCR Múltiplo (YAMAGUTI, 2008), o PCR a Tempo Real (STRAIT et al., 2008) e a amplificação de múltiplo VNTR (VRANCKX et al., 2011). A caracterização molecular das estirpes foi realizada mediante a análise do número de cópias de VNTR em P97R1, P146R3, H2R1 e H4. O M. hyopneumoniae foi detectado em amostras de suínos saudáveis e pneumônicos e a quantidade de M. hyopneumoniae nas amostras positivas variou com o tipo de ensaio. O maior número de amostras positivas foi identificado pela amplificação de múltiplas VNTR combinado com a eletroforese de capilares. Usando o PCR a Tempo Real, 4.9*10 4 cópias de genoma/mL de M. hyopneumoniae foram detectadas em pulmões aparentemente saudáveis. Uma quantidade média de 3.9*10 6 cópias de genoma/mL de M. hyopneumoniae foi detectada em pulmões pneumônicos. A análise do número de cópias de VNTR demonstrou uma elevada variabilidade genética das estirpes d...

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Section: Discussionsupporting

confidence: 52%

Section: Discussionmentioning

confidence: 99%

Detection, quantification and genetic variability of Mycoplasma hyopneumoniae from apparently healthy and pneumonic swine

Pulgarón¹,

Marques

Campos

et al. 2015

Braz. J. Vet. Res. Anim. Sci.

View full text Add to dashboard Cite

show abstract

“…2B). These regions correspond with an EKQ repeat domain with the motif ((QK))(EQ)(QK)(X) identified previously (54), which is in close proximity to the N terminus of P50 683 . The COILS2 algorithm (p Ͼ 0.9) and the Paircoil2 algorithm (p ϭ 0.0243, coiled-coil cutoff of p Ͻ 0.025) both indicated that the EKQ region forms a coiled coil.…”

Section: Mhp683mentioning

confidence: 66%

Sequence TTKF↓QE Defines the Site of Proteolytic Cleavage in Mhp683 Protein, a Novel Glycosaminoglycan and Cilium Adhesin of Mycoplasma hyopneumoniae

Bogema

Scott

Padula

et al. 2011

Journal of Biological Chemistry

141

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Mycoplasma hyopneumoniae colonizes the ciliated respiratory epithelium of swine, disrupting mucociliary function and inducing chronic inflammation. P97 and P102 family members are major surface proteins of M. hyopneumoniae and play key roles in colonizing cilia via interactions with glycosaminoglycans and mucin. The p102 paralog, mhp683, and homologs in strains from different geographic origins encode a 135-kDa preprotein (P135) that is cleaved into three fragments identified here as P45 683

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“…A number of PCR assays have been developed and reported to be both sensitive and specific for M. hyopneumoniae (4,6,11,19,20,22,24,26). As a result, this technique is now among the most widely used for detection of M. hyopneumoniae in pigs.While the development of PCR assays has greatly enhanced our ability to detect M. hyopneumoniae, the genetic variability of the organism (1,5,12,14,15,21) can affect detection by PCR. It is not known what effect this heterogeneity has on the sensitivities of the different assays.…”

mentioning

confidence: 99%

“…While the development of PCR assays has greatly enhanced our ability to detect M. hyopneumoniae, the genetic variability of the organism (1,5,12,14,15,21) can affect detection by PCR. It is not known what effect this heterogeneity has on the sensitivities of the different assays.…”

mentioning

confidence: 99%

Real-Time PCR Assays To Address Genetic Diversity among Strains ofMycoplasma hyopneumoniae

et al. 2008

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Mycoplasma hyopneumoniae is an important cause of pneumonia in pigs around the world, but confirming its presence in (or absence from) pigs can be difficult. Culture for diagnosis is impractical, and seroconversion is often delayed after natural infection, limiting the use of serology. Numerous PCR assays for the detection of M. hyopneumoniae have been developed, targeting several different genes. Recently, genetic diversity among strains of M. hyopneumoniae was demonstrated. The effect of this diversity on the accuracy and sensitivity of the M. hyopneumoniae PCR assays could result in false-negative results in current PCR tests. In this study, a panel of isolates of M. hyopneumoniae, M. flocculare, M. hyorhinis, and M. hyosynoviae were tested with a number of M. hyopneumoniae-specific PCR assays. Some M. hyopneumoniae PCR assays tested did not detect all isolates of M. hyopneumoniae. To increase the efficiency of PCR testing, two new real-time PCR assays that are specific and capable of detecting all of the M. hyopneumoniae isolates used in this study were developed.Mycoplasma hyopneumoniae is the causative agent of enzootic pneumonia and an integral component of the porcine respiratory disease complex. Considered one of the most important sources of disease-associated losses in swine production, M. hyopneumoniae is also one of the most difficult to detect. This organism is difficult to isolate in pure culture because it is easily overgrown by other contaminating bacteria; therefore, culture is generally not attempted. In addition, seroconversion to M. hyopneumoniae is often slow within a herd and can vary considerably among pigs, making the use of enzyme-linked immunosorbent assays less effective. A number of PCR assays have been developed and reported to be both sensitive and specific for M. hyopneumoniae (4,6,11,19,20,22,24,26). As a result, this technique is now among the most widely used for detection of M. hyopneumoniae in pigs.While the development of PCR assays has greatly enhanced our ability to detect M. hyopneumoniae, the genetic variability of the organism (1,5,12,14,15,21) can affect detection by PCR. It is not known what effect this heterogeneity has on the sensitivities of the different assays. Real-time PCR has many advantages over traditional PCR assays, including less time to obtain quantitative results and a decrease in environmental contamination. Together, these traits make real-time PCR assays preferred in diagnostic and research settings. A real-time assay that is based on a conserved target common among isolates of M. hyopneumoniae has not yet been described. The two currently published M. hyopneumoniae-specific real-time assays had to be used in combination in order to detect all M.hyopneumoniae field samples in a Swiss collection of isolates (6). In addition, many of the previously published PCR assays were validated using only a small number of mycoplasma isolates. Therefore, the objectives of this study were to measure the ability of several M. hyopneumoniae-specific PCR assays to de...

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Variable number of tandem aminoacid repeats in adhesion-related CDS products in Mycoplasma hyopneumoniae strains

Cited by 44 publications

References 30 publications

Detection, quantification and genetic variability of Mycoplasma hyopneumoniae from apparently healthy and pneumonic swine

Detection, quantification and genetic variability of Mycoplasma hyopneumoniae from apparently healthy and pneumonic swine

Sequence TTKF↓QE Defines the Site of Proteolytic Cleavage in Mhp683 Protein, a Novel Glycosaminoglycan and Cilium Adhesin of Mycoplasma hyopneumoniae

Real-Time PCR Assays To Address Genetic Diversity among Strains ofMycoplasma hyopneumoniae

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