实时荧光定量PCR在猪肺炎支原体检测中的应用

实时荧光定量PCR技术是一种利用荧光检测方法来定量核酸的技术,具有高度的灵敏性、特异性和精确性.综述了荧光定量PCR技术的基本原理及其在猪肺炎支原体检测中的应用.     

盘羊肺炎支原体感染PCR 检测方法的建立与初步应用

羊支原体性肺炎,是由支原体引起的一类高度接触性传染病,又称羊传染性胸膜肺炎(Infections pleuropneumonia of sheep and goats),临床症状主要表现为高热、咳嗽、消瘦、     

Microorganisms in chemotherapy for pancreatic cancer: An overview of current research and future directions

Pancreatic cancer is a malignant tumor of the digestive system with a very high mortality rate. While gemcitabine-based chemotherapy is the predominant treatment for terminal pancreatic cancer, its therapeutic effect is not satisfactory. Recently, many studies have found that microorganisms not only play a consequential role in the occurrence and progression of pancreatic cancer but also modulate the effect of chemotherapy to some extent. Moreover, microorganisms may become an important biomarker for predicting pancreatic carcinogenesis and detecting the prognosis of pancreatic cancer. However, the existing experimental literature is not sufficient or convincing. Therefore, further exploration and experiments are imperative to understanding the mechanism underlying the interaction between microorganisms and pancreatic cancer. In this review, we primarily summarize and discuss the influences of oncolytic viruses and bacteria on pancreatic cancer chemotherapy because these are the two types of microorganisms that are most often studied. We focus on some potential methods specific to these two types of microorganisms that can be used to improve the efficacy of chemotherapy in pancreatic cancer therapy.     

Detection of Mycoplasma genitalium using a wireless magnetoelastic immunosensor

A wireless immunosensor for the detection of Mycoplasma genitalium was fabricated by immobilizing polyclonal antibody onto the surface of a magnetostrictive strip. In response to a time-varying magnetic field, the immunosensor longitudinally vibrates at a resonance frequency, emitting magnetic flux that can be remotely detected by a pickup coil. No physical connections between the immunosensor and the detection system are required, facilitating wireless aseptic operation. The binding of M. genitalium to the immunosensor surface resulted in a decrease in the resonance frequency of the immunosensor. When solutions with varying concentrations of the bacteria were tested, the shift of the resonance frequency was proportional to the concentration of M. genitalium. Under the optimized conditions, the linear range for the determination of M. genitalium was 2.0 × 10³ to 2.9 × 10⁴ color change units (ccu)/ml with a detection limit of 3.4 × 10² ccu/ml. The immunosensor was successfully applied to real samples containing M. genitalium with results similar to those previously obtained by the color change unit method.     

Porcine circovirus type 2 (PCV2) vaccination is effective in reducing disease and PCV2 shedding in semen of boars concurrently infected with PCV2 and Mycoplasma hyopneumoniae

The objectives were to determine whether the amount of porcine circovirus type 2 (PCV2) shed in semen increased in boars experimentally coinfected with Mycoplasma hyopneumoniae (MHYO), and whether PCV2 vaccination of boars prior to PCV2 exposure reduced PCV2 viremia and virus shedding in semen. Twelve specific-pathogen-free PCV2- and MHYO-naïve boars were randomly and equally assigned to one of four groups. Six boars were vaccinated against PCV2 (VAC) on Day 0; three PCV2 vaccinated and three non-vaccinated boars were inoculated with MHYO on Day 21, and all boars were challenged with PCV2 on Day 35. The four treatment groups included PCV2-Infected (I), VAC-PCV2-I, MHYO-PCV2-Coinfected (CoI), and VAC-MHYO-PCV2-CoI. Semen, blood swabs, feces, and serum samples were collected weekly until Day 70. All vaccinated boars had seroconverted to PCV2 by Day 35. Between Days 28 and 35, MHYO boars developed moderate respiratory disease, characterized by coughing, respiratory distress, mucopurulent nasal discharge and loss of body condition. One MHYO-PCV2-CoI boar died on Day 50. Boars in the PCV2-I and MHYO-PCV2-CoI groups had significantly higher PCV2 DNA loads in blood swabs than the remaining boars. Moreover, PCV2 vaccination significantly reduced the incidence and amount of PCV2 shedding in semen and feces. In summary, although concurrent MHYO infection did not influence PCV2 shedding patterns, coinfection of boars with PCV2 and MHYO resulted in severe clinical disease and viral shedding was significantly decreased by PCV2 vaccination.     

Mycoplasma pneumoniae infection induces a neutrophil-derived antimicrobial peptide, cathelin-related antimicrobial peptide

In innate immunity, cationic antimicrobial peptides including cathelin-related antimicrobial peptide (CRAMP) are known to play critical roles in protecting the host from infection by invasive microbes, including Gram-positive and -negative bacteria. However, little is known about the interactions between CRAMP and mycoplasmas. In the present study, the antimicrobial activity of CRAMP against M. pneumoniae and the expression of CRAMP in bronchoalveolar lavage fluid (BALF) of M. pneumoniae-infected mice was examined. CRAMP at 10-20 μg/mL reduced the growth of two strains of M. pneumoniae by 100 to 1000-fold. The amount of CRAMP in the BALF of M. pneumoniae-infected mice was 20～25 ng/mL by ELISA. The presence of mature CRAMP in BALF was observed by Western blotting. Neutrophils in BALF showed a fair amount of CRAMP in their cytoplasm by immunofluorescence. Furthermore, the addition of M. pneumoniae resulted in the release of a large amount of CRAMP from neutrophils induced by thioglycolate. These results suggest that CRAMP from neutrophils may play an important role in protection against M. pneumoniae infection.     

Expression and characterization of a Mycoplasma genitalium glycosyltransferase in membrane glycolipid biosynthesis: potential target against mycoplasma infections

Mycoplasmas contain glycoglycerolipids in their plasma membrane as key structural components involved in bilayer properties and stability. A membrane-associated glycosyltransferase (GT), GT MG517, has been identified in Mycoplasma genitalium, which sequentially produces monoglycosyl- and diglycosyldiacylglycerols. When recombinantly expressed in Escherichia coli, the enzyme was functional in vivo and yielded membrane glycolipids from which Glcβ1,6GlcβDAG was identified as the main product. A chaperone co-expression system and extraction with CHAPS detergent afforded soluble protein that was purified by affinity chromatography. GT MG517 transfers glucosyl and galactosyl residues from UDP-Glc and UDP-Gal to dioleoylglycerol (DOG) acceptor to form the corresponding β-glycosyl-DOG, which then acts as acceptor to give β-diglycosyl-DOG products. The enzyme (GT2 family) follows Michaelis-Menten kinetics. k(cat) is about 5-fold higher for UDP-Gal with either DOG or monoglucosyldioleoylglycerol acceptors, but it shows better binding for UDP-Glc than UDP-Gal, as reflected by the lower K(m), which results in similar k(cat)/K(m) values for both donors. Although sequentially adding glycosyl residues with β-1,6 connectivity, the first glycosyltransferase activity (to DOG) is about 1 order of magnitude higher than the second (to monoglucosyldioleoylglycerol). Because the ratio between the non-bilayer-forming monoglycosyldiacylglycerols and the bilayer-prone diglycosyldiacylglycerols contributes to regulate the properties of the plasma membrane, both synthase activities are probably regulated. Dioleoylphosphatidylglycerol (anionic phospholipid) activates the enzyme, k(cat) linearly increasing with dioleoylphosphatidylglycerol concentration. GT MG517 is shown to be encoded by an essential gene, and the addition of GT inhibitors results in cell growth inhibition. It is proposed that glycolipid synthases are potential targets for drug discovery against infections by mycoplasmas.     

Comparative genomics identifies genes shared by distantly related insect-transmitted plant pathogenic mollicutes

Phytoplasmas and spiroplasmas are distantly related insect-transmitted plant pathogens within the class Mollicutes. Genome sequencing projects of phytoplasma strain Aster Yellows-Witches' Broom (AY-WB) and Spiroplasma kunkelii are near completion. Complete genome sequences of seven obligate animal and human pathogenic mollicutes (Mycoplasma and Ureaplasma spp.), and OY phytoplasma have been reported. Putative ORFs predicted from the genome sequences of AY-WB and S. kunkelii were compared to those of the completed genomes. This resulted in identification of at least three ORFs present in AY-WB, OY and S. kunkelii but not in the obligate animal and human pathogenic mollicutes. Moreover, we identified ORFs that seemed more closely related between AY-WB and S. kunkelii than to their mycoplasma counterparts. Phylogenetic analyses using parsimony were employed to study the origin of these genes, resulting in identification of one gene that may have undergone horizontal gene transfer. The possible involvement of these genes in plant pathogenicity is discussed.     

支原体肺炎患儿血清中相关炎性因子的表达变化情况及其临床意义探讨

目的：探讨支原体肺炎（Mycoplasma Pneumoniae Pneumonia,MPP）患儿血清中细胞因子IL-8,IL-12的表达水平及hs-CRP、IgG和血清补体（C）的变化及其临床意义。方法：收集MPP患儿50例,分为重症组、轻症组。健康儿童42例作为对照组;用ELISA法测定MPP患儿急性期、恢复期及对照组儿童血清IL-8、IL-12的水平,用血浆蛋白分析仪速率散射比浊法测定hs-CRP,Ig和C含量。结果：在急性期和恢复期MPP患儿血清IL-12含量明显低于正常对照组（P〈0.05）;而血清IL-8含量在急性期明显高于正常对照组（P〈0.01）。重症组患儿血清中IL-12明显低于轻症组,而血清中IL-8较轻症组高（P均〈0.01）。急性期MPP患儿血清IgM,IgG与对照组相比明显升高（P均〈0.01）;而IgA明显降低（P〈0.05）。急性期MPP患儿hs-CRP、C3、C4与对照组比较显著升高（分别为P〈0.01、P〈0.01、P〈0.05）。重症组患儿血清中IgM,IgG与轻症组相比明显升高（P均〈0.01）;IgA与轻症组相比明显降低（P〈0.05）;重症组患儿血清中hs-CRP、C3、C4与轻症组相比明显升高（P均〈0.01）。结论：检测相关血清炎性细胞因子对判定MPP患儿的病情和预后有较高的临床应用价值。