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1.
Orthopadie (Heidelb) ; 53(4): 297-307, 2024 Apr.
Article in German | MEDLINE | ID: mdl-38376535

ABSTRACT

The posterolateral corner of the knee is composed of a multilayered complex of ligamentous, musculotendinous, and capsular structures, which interact in a synergistic stabilizing manner with the central structures, particularly the posterior cruciate ligament. Injuries of the posterolateral corner are generally accompanied by rupture of the posterior cruciate ligament. Depending on the injured structures, injuries of the posterolateral corner result in posterolateral rotational instability alone (Fanelli A) or with lateral instability (Fanelli B/C). For rotational instability alone, isolated popliteus bypass is an effective procedure; with concomitant lateral instability in flexion, additional stabilization of the lateral collateral ligament is required. Most of the various available techniques are described as open reconstruction procedures. In recent years, arthroscopic techniques for posterolateral reconstruction have also been successfully developed.


Subject(s)
Joint Instability , Knee Injuries , Posterior Cruciate Ligament , Humans , Joint Instability/surgery , Knee Joint/surgery , Posterior Cruciate Ligament/surgery , Knee , Knee Injuries/surgery
3.
Unfallchirurgie (Heidelb) ; 126(1): 55-66, 2023 Jan.
Article in German | MEDLINE | ID: mdl-36542118

ABSTRACT

The treatment of extra-articular proximal tibial fractures is a therapeutic challenge due to the frequently significant soft tissue injury, the effect of the deforming forces and the need for an exact restoration of the bony alignment. Various methods of osteosynthesis are available for surgical stabilization. The locking plate osteosynthesis is the most frequently used procedure because of its good biomechanical stability, especially in osteoporotic bones, and the protection of the periosteal blood flow. Depending on the extent and stability of the defect zone, especially in the case of a medial comminuted zone and the bone quality, bilateral plate osteosynthesis can be necessary. If the proximal fragment is big enough, closed reduction and intramedullary nailing are possible. In the case of severely compromised soft tissue or very short epiphyseal fragments, the construction of an external fixator, e.g. hybrid external fixator, is recommended, which also allows definitive treatment under early full weight bearing. The most important complications are axial and torsional malalignments.


Subject(s)
Fracture Fixation, Intramedullary , Fractures, Comminuted , Tibial Fractures , Humans , Tibial Fractures/diagnostic imaging , Fracture Fixation, Internal/methods , Tibia , Fracture Fixation, Intramedullary/methods , Fractures, Comminuted/diagnostic imaging
4.
Eur J Trauma Emerg Surg ; 49(3): 1433-1439, 2023 Jun.
Article in English | MEDLINE | ID: mdl-36484798

ABSTRACT

INTRODUCTION: The objective of this investigation was to compare different techniques to improve visualization and reduction in tibial plateau fractures involving the central lateral segments. METHODS: Matched pairs of pre-fractured cadaveric tibial plateau fractures that include the central lateral segments were treated by either an anterolateral approach (supine) or PL approach (prone). Reduction was stepwise extended by additional fracturoscopy (FS), nanoscopy (NS) and lastly by epicondyle osteotomy (ECO). Reduction was analyzed by 3D scan and visualization of the lateral plateau was quantified. RESULTS: Ten specimens (3 pairs 41B3.1, 2 pairs 41C3.3) were analyzed. Fracture steps involving the antero-latero-central (ALC) segment were insufficiently reduced after fluoroscopy using both approaches (AL 2.2 ± 1.2 mm vs PL 2.2 ± 1.0 mm, p 0.95). Additional NS and ECO achieved optimized fracture reduction in the ALC segment (NS AL 1.6 ± 1.3 mm vs PL 0.8 ± 0.9 mm, p 0.32). NS provided visualization of the entire lateral plateau (PL 102.9% ± 7.4, AL 108.8 ± 19.2%), while fracturoscopy only allowed visualization of the ALL segment and partially of PLL and ALC segments (PL 22.0 ± 23.4%, AL 29.7 ± 18.3%). CONCLUSION: Optimized reduction of tibial head fractures with involvement of latero-central segments requires additional video-assisted reduction or extended approaches. Nanoscopy helps visualizing of the entire lateral plateau, when compared to fracturoscopy and may become a valuable reduction aid.


Subject(s)
Tibial Fractures , Tibial Plateau Fractures , Humans , Fracture Fixation, Internal/methods , Tibial Fractures/diagnostic imaging , Tibial Fractures/surgery , Tibia/diagnostic imaging , Tibia/surgery , Cadaver , Treatment Outcome
5.
Medicine (Baltimore) ; 99(42): e22412, 2020 Oct 16.
Article in English | MEDLINE | ID: mdl-33080675

ABSTRACT

In many German trauma centres, it is routine to perform abdominal follow-up sonography (AFS) 6 h after admission for patients with multiple trauma, even if the clinical course is uneventful and multi-slice computed tomography (MSCT) reveals no abdominal pathology. However, this approach is not recommended in the German Guidelines for trauma, and recent studies have questioned the value of AFS to these patients. The present study aimed to evaluate the revised German Guidelines for trauma with respect to the omission of AFS.We included patients with multiple injuries with no clinical signs of abdominal trauma and with normal abdominal MSCT. We collected clinical data of 370 consecutive patients who underwent AFS (Group A) and another 370 consecutive patients who did not undergo AFS (Group B).No abdominal injury was missed by the omission of AFS, and thus, no patient suffered from its omission or benefitted from the use of AFS. In our study population, the negative predictive value of normal MSCT results combined with no clinical signs of abdominal trauma was 100% (95% confidence interval: 99.5%-100.0%).This single-centre study conducted in a large German trauma centre demonstrates AFS to have no utility in the diagnosis of abdominal injury. Moreover, omission of AFS for conscious patients without clinical signs of abdominal trauma and with negative abdominal MSCT does not appear to have negative consequences in terms of missed abdominal injury.Therefore, AFS can be safely omitted in the majority of cases of polytrauma, which simplifies the imaging workup tremendously.


Subject(s)
Abdominal Injuries/diagnostic imaging , Multiple Trauma/diagnostic imaging , Practice Guidelines as Topic , Ultrasonography/methods , Adolescent , Adult , Aged , Aged, 80 and over , Child , Child, Preschool , Female , Germany , Humans , Infant , Male , Middle Aged , Retrospective Studies , Trauma Centers , Trauma Severity Indices , Unnecessary Procedures
6.
BMC Musculoskelet Disord ; 19(1): 232, 2018 Jul 18.
Article in English | MEDLINE | ID: mdl-30021587

ABSTRACT

BACKGROUND: Postoperative pain control and enhanced mobilization, muscle strength and range of motion following total knee arthroplasty (TKA) are pivotal requisites to optimize rehabilitation and early recovery. The aim of the study was to analyze the effect of local infiltration analgesia (LIA), peripheral nerve blocks, general and spinal anesthesia on early functional recovery and pain control in primary total knee arthroplasty. METHODS: Between January 2016 until August 2016, 280 patients underwent primary TKA and were subdivided into four groups according to their concomitant pain and anesthetic procedure with catheter-based techniques of femoral and sciatic nerve block (group GA&FNB, n = 81) or epidural catheter (group SP&EPI, n = 51) in combination with general anesthesia or spinal anesthesia, respectively, and LIA combined with general anesthesia (group GA&LIA, n = 86) or spinal anesthesia (group SP&LIA, n = 61). Outcome parameters focused on the evaluation of pain (NRS scores), mobilization, muscle strength and range of motion up to 7 days postoperatively. The cumulative consumption of (rescue) pain medication was analyzed. RESULTS: Pain relief was similar in all groups, while the use of opioid medication was significantly lower (up to 58%) in combination with spinal anesthesia, especially in SP&EPI. The LIA groups, in contrast, revealed significant higher mobilization (up to 26%) and muscle strength (up to 20%) in the early postoperative period. No analgesic technique-related or surgery-related complications occurred within the first 7 days. Due to insufficient pain relief, 8.4% of the patients in the catheter-based groups and 12.2% in the LIA groups resulted in a change of the anesthetics pain management. CONCLUSIONS: The LIA technique offers a safe and effective treatment option concerning early functional recovery and pain control in TKA. Significant advantages were shown for mobilization and muscle strength in the early postoperative period while pain relief was comparable within the groups.


Subject(s)
Anesthesia, General/trends , Anesthesia, Local/trends , Anesthesia, Spinal/trends , Arthroplasty, Replacement, Knee/trends , Autonomic Nerve Block/trends , Pain Management/trends , Aged , Aged, 80 and over , Anesthesia, General/methods , Anesthesia, Local/methods , Anesthesia, Spinal/methods , Arthroplasty, Replacement, Knee/methods , Autonomic Nerve Block/methods , Female , Humans , Male , Middle Aged , Pain Management/methods , Pain, Postoperative/diagnostic imaging , Pain, Postoperative/prevention & control , Range of Motion, Articular/drug effects , Range of Motion, Articular/physiology , Recovery of Function/drug effects , Recovery of Function/physiology , Retrospective Studies , Treatment Outcome
7.
BMC Musculoskelet Disord ; 19(1): 249, 2018 Jul 24.
Article in English | MEDLINE | ID: mdl-30037342

ABSTRACT

BACKGROUND: The aim of the study was to analyze the effect of local infiltration analgesia (LIA), peripheral nerve blocks, general and spinal anesthesia on early functional recovery and pain control in primary unicompartmental knee arthroplasty (UKA). METHODS: Between January 2016 until August 2016, 134 patients underwent primary UKA and were subdivided into four groups according to their concomitant pain and anesthetic procedure with catheter-based techniques of femoral and sciatic nerve block (group GA&FNB, n = 38) or epidural catheter (group SP&EPI, n = 20) in combination with general anesthesia or spinal anesthesia, respectively, and LIA combined with general anesthesia (group GA&LIA, n = 46) or spinal anesthesia (group SP&LIA, n = 30). Outcome parameters focused on the evaluation of pain (NRS scores), mobilization, muscle strength and range of motion up to 7 days postoperatively. The cumulative consumption of (rescue) pain medication was analyzed. RESULTS: The LIA groups revealed significantly lower (about 50%) mean NRS scores (at rest) compared to the catheter-based groups at the day of surgery. In the early postoperative period, the dose of hydromorphone as rescue pain medication was significantly lower (up to 68%) in patients with SP&EPI compared to all other groups. No significant differences could be detected with regard to grade of mobilization, muscle strength and range of motion. However, there seemed to be a trend towards improved mobilization and muscle strength with general anesthesia and LIA, whereof general anesthesia generally tended to ameliorate mobilization. CONCLUSIONS: Except for a significant lower NRS score at rest in the LIA groups at day of surgery, pain relief was comparable in all groups without clinically relevant differences, while the use of opioids was significantly lower in patients with SP&EPI. A clear clinically relevant benefit for LIA in UKA cannot be stated. However, LIA offers a safe and effective treatment option comparable to the well-established conventional procedures.


Subject(s)
Anesthesia, General/trends , Anesthesia, Local/trends , Anesthesia, Spinal/trends , Arthroplasty, Replacement, Knee/trends , Autonomic Nerve Block/trends , Recovery of Function/physiology , Aged , Aged, 80 and over , Anesthesia, General/methods , Anesthesia, Local/methods , Anesthesia, Spinal/methods , Anesthetics, Local/administration & dosage , Arthroplasty, Replacement, Knee/adverse effects , Arthroplasty, Replacement, Knee/methods , Autonomic Nerve Block/methods , Female , Humans , Male , Middle Aged , Osteoarthritis, Knee/diagnosis , Osteoarthritis, Knee/surgery , Pain Management/methods , Pain Management/trends , Pain, Postoperative/etiology , Pain, Postoperative/prevention & control , Recovery of Function/drug effects , Retrospective Studies , Treatment Outcome
8.
Unfallchirurg ; 121(4): 335-338, 2018 Apr.
Article in German | MEDLINE | ID: mdl-29500508

ABSTRACT

This article describes the operative stabilization of a flail chest due to traumatic serial rib fractures with extensive chest wall deformation and respiratory insufficiency. Initial conservative treatment including systemic and regional pain management and non-invasive positive pressure ventilation did not improve the pain or ventilation. Therefore, a single-port video-assisted thoracoscopic surgery (VATS) assisted internal fixation of the ribs was performed. The thoracoscopy enabled easy repositioning of the ribs and additionally an estimation of intrathoracic injuries.


Subject(s)
Accidental Falls , Fracture Fixation, Internal/methods , Fractures, Multiple/surgery , Minimally Invasive Surgical Procedures/methods , Rib Fractures/surgery , Thoracic Surgery, Video-Assisted/methods , Aged , Female , Fractures, Multiple/diagnostic imaging , Hemothorax/diagnostic imaging , Hemothorax/surgery , Humans , Imaging, Three-Dimensional , Postoperative Care/methods , Rib Fractures/diagnostic imaging , Tomography, X-Ray Computed
9.
Transbound Emerg Dis ; 61(6): 500-10, 2014 Dec.
Article in English | MEDLINE | ID: mdl-23347819

ABSTRACT

The epidemiological situation of foot-and-mouth disease virus (FMDV) is uncertain in Nigeria, where the disease is endemic, and the majority of outbreaks are unreported. Control measures for FMD in Nigeria are not being implemented due to the absence of locally produced vaccines and an official ban on vaccine importation. This study summarizes the findings of a 3-year study aimed at quantifying the seroprevalence of FMD, its distribution in susceptible species and the genetic diversity of FMDV isolated from the Plateau State of Nigeria. A 29% FMD prevalence was estimated using 3ABC enzyme-linked immunosorbent assay (3ABC ELISA). Farms with suspected FMD nearby, with contact with wildlife, that used drugs or FMD vaccines or with >100 animals, and animals of large ruminant species and in pastures other than nomadic grazing were significantly (P < 0.05) associated with FMD. Antibodies against five FMDV serotypes, (A, O, SAT1, SAT2 and SAT3) were detected by the virus neutralization test (VNT) at various titres (<100->800) from all tested sera from most parts of the region. This is probably the first report of the presence of FMDV SAT3 in Nigeria. Further studies to investigate the potential probable presence and prevalence of SAT 3 virus in Nigeria are required. Tissue samples collected from clinical animals were positive for FMDV. Virus isolates were sequenced and confirmed as serotype A. All of the isolates showed marked genetic homogeneity with >99% genetic identity in the VP1 region and were most closely related to a previously described virus collected from Cameroon in 2000. This study provides knowledge on the epidemiological situation of FMD in Plateau State, Nigeria, and will probably help to develop effective control and preventive strategies for the disease in Nigeria and other countries in the West African subregion.


Subject(s)
Cattle Diseases , Foot-and-Mouth Disease Virus/immunology , Foot-and-Mouth Disease/epidemiology , Animals , Antibodies, Viral/blood , Cattle , Cattle Diseases/blood , Cattle Diseases/epidemiology , Cattle Diseases/virology , Enzyme-Linked Immunosorbent Assay/veterinary , Foot-and-Mouth Disease/blood , Foot-and-Mouth Disease Virus/isolation & purification , Genotype , Neutralization Tests/veterinary , Nigeria/epidemiology , Prevalence , Seroepidemiologic Studies
10.
Phys Rev Lett ; 107(12): 120401, 2011 Sep 16.
Article in English | MEDLINE | ID: mdl-22026757

ABSTRACT

We report on the observation of triatomic Efimov resonances in an ultracold gas of cesium atoms. Exploiting the wide tunability of interactions resulting from three broad Feshbach resonances in the same spin channel, we measure magnetic-field dependent three-body recombination loss. The positions of the loss resonances yield corresponding values for the three-body parameter, which in universal few-body physics is required to describe three-body phenomena and, in particular, to fix the spectrum of Efimov states. Our observations show a robust universal behavior with a three-body parameter that stays essentially constant.

11.
Transbound Emerg Dis ; 58(4): 358-71, 2011 Aug.
Article in English | MEDLINE | ID: mdl-21418546

ABSTRACT

Experimental studies of foot-and-mouth disease (FMD) in feral swine are limited, and data for clinical manifestations and disease transmissibility are lacking. In this report, feral and domestic swine were experimentally infected with FMDV (A24-Cruzeiro), and susceptibility and virus transmission were studied. Feral swine were proved to be highly susceptible to A-24 Cruzeiro FMD virus by intradermal inoculation and by contact with infected domestic and feral swine. Typical clinical signs in feral swine included transient fever, lameness and vesicular lesions in the coronary bands, heel bulbs, tip of the tongue and snout. Domestic swine exhibited clinical signs of the disease within 24 h after contact with feral swine, whereas feral swine did not show clinical signs of FMD until 48 h after contact with infected domestic and feral swine. Clinical scores of feral and domestic swine were comparable. However, feral swine exhibited a higher tolerance for the disease, and their thicker, darker skin made vesicular lesions difficult to detect. Virus titration of oral swabs showed that both feral and domestic swine shed similar amounts of virus, with levels peaking between 2 to 4 dpi/dpc (days post-inoculation/days post-contact). FMDV RNA was intermittently detectable in the oral swabs by real-time RT-PCR of both feral and domestic swine between 1 and 8 dpi/dpc and in some instances until 14 dpi/12 dpc. Both feral and domestic swine seroconverted 6-8 dpi/dpc as measured by 3ABC antibody ELISA and VIAA assays. FMDV RNA levels in animal room air filters were similar in feral and domestic swine animal rooms, and were last detected at 22 dpi, while none were detectable at 28 or 35 dpi. The FMDV RNA persisted in domestic and feral swine tonsils up to 33-36 dpi/dpc, whereas virus isolation was negative. Results from this study will help understand the role feral swine may play in sustaining an FMD outbreak, and may be utilized in guiding surveillance, epidemiologic and economic models.


Subject(s)
Foot-and-Mouth Disease/transmission , Swine Diseases/epidemiology , Air Microbiology , Animals , Animals, Wild , Disease Susceptibility , Female , Foot-and-Mouth Disease/pathology , Foot-and-Mouth Disease/virology , Foot-and-Mouth Disease Virus/isolation & purification , Male , Swine , Swine Diseases/pathology , Swine Diseases/virology , Time Factors
12.
Transbound Emerg Dis ; 57(5): 315-29, 2010 Oct.
Article in English | MEDLINE | ID: mdl-20629970

ABSTRACT

In 2007, Vietnam experienced swine disease outbreaks causing clinical signs similar to the 'porcine high fever disease' that occurred in China during 2006. Analysis of diagnostic samples from the disease outbreaks in Vietnam identified porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV-2). Additionally, Escherichia coli and Streptococcus equi subspecies zooepidemicus were cultured from lung and spleen, and Streptococcus suis from one spleen sample. Genetic characterization of the Vietnamese PRRSV isolates revealed that this virus belongs to the North American genotype (type 2) with a high nucleotide identity to the recently reported Chinese strains. Amino acid sequence in the nsp2 region revealed 95.7-99.4% identity to Chinese strain HUN4, 68-69% identity to strain VR-2332 and 58-59% identity to strain MN184. A partial deletion in the nsp2 gene was detected; however, this deletion did not appear to enhance the virus pathogenicity in the inoculated pigs. Animal inoculation studies were conducted to determine the pathogenicity of PRRSV and to identify other possible agents present in the original specimens. Pigs inoculated with PRRSV alone and their contacts showed persistent fever, and two of five pigs developed cough, neurological signs and swollen joints. Necropsy examination showed mild to moderate bronchopneumonia, enlarged lymph nodes, fibrinous pericarditis and polyarthritis. PRRSV was re-isolated from blood and tissues of the inoculated and contact pigs. Pigs inoculated with lung and spleen tissue homogenates from sick pigs from Vietnam developed high fever, septicaemia, and died acutely within 72 h, while their contact pigs showed no clinical signs throughout the experiment. Streptococcus equi subspecies zooepidemicus was cultured, and PRRSV was re-isolated only from the inoculated pigs. Results suggest that the cause of the swine deaths in Vietnam is a multifactorial syndrome with PRRSV as a major factor.


Subject(s)
Communicable Diseases, Emerging/veterinary , Porcine Reproductive and Respiratory Syndrome/virology , Porcine respiratory and reproductive syndrome virus/genetics , Porcine respiratory and reproductive syndrome virus/pathogenicity , Animals , Arthritis/pathology , Communicable Diseases, Emerging/epidemiology , Disease Outbreaks/veterinary , Lymph Nodes/pathology , Pericardium/pathology , Phylogeny , Pneumonia/epidemiology , Porcine Reproductive and Respiratory Syndrome/epidemiology , Porcine Reproductive and Respiratory Syndrome/pathology , Swine , Vietnam/epidemiology
13.
Phys Rev Lett ; 104(5): 053201, 2010 Feb 05.
Article in English | MEDLINE | ID: mdl-20366759

ABSTRACT

We report on the observation of an elementary exchange process in an optically trapped ultracold sample of atoms and Feshbach molecules. We can magnetically control the energetic nature of the process and tune it from endoergic to exoergic, enabling the observation of a pronounced threshold behavior. In contrast to relaxation to more deeply bound molecular states, the exchange process does not lead to trap loss. We find excellent agreement between our experimental observations and calculations based on the solutions of three-body Schrödinger equation in the adiabatic hyperspherical representation. The high efficiency of the exchange process is explained by the halo character of both the initial and final molecular states.

14.
Phys Rev Lett ; 102(14): 140401, 2009 Apr 10.
Article in English | MEDLINE | ID: mdl-19392415

ABSTRACT

We report on the measurement of four-body recombination rate coefficients in an atomic gas. Our results obtained with an ultracold sample of cesium atoms at negative scattering lengths show a resonant enhancement of losses and provide strong evidence for the existence of a pair of four-body states, which is strictly connected to Efimov trimers via universal relations. Our findings confirm recent theoretical predictions and demonstrate the enrichment of the Efimov scenario when a fourth particle is added to the generic three-body problem.

15.
Phys Rev Lett ; 101(2): 023201, 2008 Jul 11.
Article in English | MEDLINE | ID: mdl-18764179

ABSTRACT

We study inelastic collisions in a pure, trapped sample of Feshbach molecules made of bosonic cesium atoms in the quantum halo regime. We measure the relaxation rate coefficient for decay to lower-lying molecular states and study the dependence on scattering length and temperature. We identify a pronounced loss minimum with varying scattering length along with a further suppression of loss with decreasing temperature. Our observations provide insight into the physics of a few-body quantum system that consists of four identical bosons at large values of the two-body scattering length.

16.
Vet Microbiol ; 47(1-2): 157-65, 1995 Nov.
Article in English | MEDLINE | ID: mdl-8604547

ABSTRACT

An acute, fatal disease of rabbits, termed Rabbit Hemorrhagic Disease (RHD), has occurred in Asia, Europe, and North America since 1984. The clinical signs, pathologic lesions, and epidemiologic features seen in the various outbreaks were very similar. Although RHD virus (RHDV) was initially characterized as a picornavirus or a parvovirus, it is now proven to be a calicivirus. This study compared the immune responses generated following the vaccination and challenge inoculation of rabbits with isolates of RHDV obtained from Italy, Korea, Mexico, and Spain. The cross-reactivity of antisera was demonstrated using the hemagglutination inhibition (HI) test. There were minimal differences between the homologous and heterologous responses. Statistical analysis of the HI results showed no difference between the isolates within the Italian, Korean, and Spanish vaccinate groups. The difference obtained between the Mexican and Korean antigens within the Mexican vaccinate group is likely due to individual animal variation. The similarity of the isolates was also demonstrated using a monoclonal antibody directed against RHDV.


Subject(s)
Antibodies, Viral/immunology , Caliciviridae Infections/veterinary , Hemorrhagic Disease Virus, Rabbit/immunology , Rabbits , Animals , Antibodies, Viral/biosynthesis , Antigens, Viral/immunology , Caliciviridae Infections/immunology , Caliciviridae Infections/virology , Cross Reactions , Female , Hemagglutination Inhibition Tests/veterinary , Immune Sera/immunology , Italy , Korea , Male , Mexico , Spain , Vaccination/veterinary , Viral Vaccines/immunology
18.
J Clin Microbiol ; 31(2): 428-32, 1993 Feb.
Article in English | MEDLINE | ID: mdl-8432831

ABSTRACT

Gamma irradiation effectively inactivated gradient-purified rinderpest virus. Irradiated antigen and sera remained functional in enzyme-linked immunosorbent assays, virus neutralization tests, and indirect fluorescent-antibody tests. Irradiation, however, led to a dose-dependent decrease in reactivity, particularly significant (P < 0.05) when both reagents were irradiated. To avoid false-positive reactions, only one reagent (serum or antigen) may be irradiated.


Subject(s)
Antigens, Viral/radiation effects , Rinderpest virus/immunology , Animals , Antibodies, Viral/radiation effects , Cattle , Enzyme-Linked Immunosorbent Assay , Fluorescent Antibody Technique , Gamma Rays , Neutralization Tests , Rinderpest/diagnosis , Rinderpest virus/radiation effects , Virology/methods
19.
Anal Biochem ; 206(1): 91-7, 1992 Oct.
Article in English | MEDLINE | ID: mdl-1456447

ABSTRACT

A simple and rapid method for cloning of amplification products directly from the polymerase chain reaction (PCR) has been developed. The method is based on the addition of a 12-base dUMP-containing sequence (CUACUACUACUA) to the 5' end of PCR primers. Incorporation of these primers during PCR results in the selective placement of dUMP residues into the 5' end of amplification products. Selective degradation of the dUMP residues in the PCR products with uracil DNA glycosylase (UDG) disrupts base pairing at the termini and generates 3' overhangs. Annealing of 3' protruding termini to vector DNA containing complementary 3' ends results in chimeric molecules which can be transformed, with high efficiency, without in vitro ligation. Directional cloning of PCR products has also been accomplished by incorporating different dU-containing sequences at the end of each PCR primer. Substitution of all dT residues in PCR primers with dU eliminates cloning of aberrant "primer dimer" products and enriches cloning of genuine PCR products. The method has been applied to cloning of inter-Alu DNA sequences from human placental DNA. Using a single primer, DNA sequences between appropriately oriented Alu sequences were amplified and cloned. Cloning of cDNA for the glyceraldehyde-3'-phosphate dehydrogenase gene from rat brain RNA was also demonstrated. The 3' end region of this gene was amplified by the 3' RACE method and the amplified DNA was cloned after UDG digestion. Characterization of cloned DNAs by sequence analysis showed accurate repair of the cloning junctions. The ligase-free cloning method with UDG should prove to be a widely applicable procedure for rapid cloning of PCR-amplified DNA.


Subject(s)
DNA Glycosylases , DNA/genetics , N-Glycosyl Hydrolases , Polymerase Chain Reaction/methods , Animals , Base Sequence , Cloning, Molecular , Deoxyuracil Nucleotides , Gene Amplification , Genome , Glyceraldehyde-3-Phosphate Dehydrogenases/genetics , Molecular Sequence Data , Rats , Repetitive Sequences, Nucleic Acid , Uracil-DNA Glycosidase
20.
Science ; 253(5023): 952, 1991 Aug 30.
Article in English | MEDLINE | ID: mdl-1887224
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