Effect of enoxacin on colonic microflora of healthy volunteers

Ten healthy volunteers received 400 mg enoxacin orally twice daily for seven days. Fecal specimens were collected at 0, 2, 4, 7, 9, 11, 16 and 21 days to study the effect of enoxacin on the normal colonic microflora. On the seventh day the fecal mean concentration of enoxacin was 348 mg/kg feces. Whereas enterobacteria were strongly suppressed during the administration of enoxacin, the gram-positive and anaerobic microflora was not significantly altered. Two weeks after enoxacin was discontinued, the colonic microflora had returned to normal.     

Effects of delayed culture on semiquantitative urinary bacteriology results.

Urine specimens received in the laboratory were cultured at intervals of two hours on three successive occasions. During the intervening period the specimens were left at room temperature. After 24 hours' incubation counts were done, and it was observed that the significance of bacterial growth was not altered by delay in culture.     

Effect of delay in processing on lysis-centrifugation blood culture results from marrow transplant patients.

The effect of delay in processing on results of lysis-centrifugation (LC; Isolator) blood cultures was assessed in 4,577 paired blood specimens. Blood specimens were obtained at all hours from 384 febrile marrow transplant patients with indwelling venous catheters and were processed by the LC technique and by a conventional two-bottle method. Most patients (84%) were receiving broad-spectrum antibiotics at the time of blood culture. Specimens were delivered to the laboratory, where Isolator tubes were held at 35 degrees C and processed in batches between 0700 and 1730 h daily. This procedure resulted in a delay beyond the manufacturer-suggested processing time of less than 8 h for 1,853 (42%) of the LC cultures. There was no overall difference in the recovery of organisms present in LC cultures processed after being held for 8 to 24 h compared with the conventional two-bottle method. LC methodology had shorter time to detection than the conventional method for detection of Candida spp. and Pseudomonas spp. (P less than 0.05). However, time to detection for Streptococcus spp. and members of the family Enterobacteriaceae, responsible for 16.3% of total isolates, was prolonged significantly by delay in processing when compared with the conventional two-bottle method (P less than 0.01). Results of this study support the recommendation of the manufacturer for processing of Isolator tubes within 8 h or less. Although one can safely delay processing beyond 8 h in terms of total recovery of organisms, such delays were associated with longer time to detection for certain important potentially pathogenic organisms which accounted for a sizeable proportion of blood culture isolates from marrow transplant patients.     

Lack of effect of orally administered human serum immunoglobulin on the normal human oral and intestinal microflora

The aim of this study was to investigate the influence of large doses of orally administered human IgG on the normal gastrointestinal microflora of healthy volunteers since human immunoglobulin has been tried as oral prophylaxis and therapy in gastrointestinal infections. Ten adult healthy volunteers received 10 g of IgG orally, once daily for three consecutive days. Aerobic and anaerobic microorganisms were identified in the saliva and stool specimens, using morphological, biochemical and serological tests and gas-liquid chromatography. Although the immunoglobulin preparation contains antibodies against a variety of microorganisms, there were no significant changes in the numbers of different aerobic and anaerobic microorgansims due to the oral intake of the immunoglobulin. IgG may, therefore, be used against pathogens without disturbing the normal oral and intestinal microflora.     

Effects of time lapse between sputum collection and culturing on isolation of clinically significant fungi.

A retrospective study of 34,314 sputum specimens received by a reference laboratory over a 10-year period demonstrated that clinically significant fungi could be isolated even after long periods of delay between collecting and culturing. As a result of this study, it should be stressed that although immediate culturing for fungi is the ideal, specimens should not be rejected because of delays in transport.     

病理报告延迟相关因素分析

目的 分析病理报告延迟的相关因素,为病理科服务质量改进提供参考途径.方法 从浙江省东阳市人民医院1999-2006年8年的病理报告中,以月为单位整群随机抽取24个月共21 038份病理报告,按活检标本、手术标本和冷冻切片标本分别统计报告的及时性,分析延迟报告发生的原因.结果 及时报告19 579份,及时报告率93.06%,延迟报告1459份,延迟报告率6.94%,其中活检报告延迟率6.02%(665/11 052),手术标本报告延迟率7.26%(643/8858),冷冻报告延迟率13.39%(151/1128).延迟报告的原因,以技术原因为主,占1158份(79.37%),其中免疫组织化学或特殊染色、科内讨论或院外会诊、访视患者或与临床沟通比例较高.责任原因占301份(20.63%),其中申请单缺项和缺既往病理资料最多见.发生环节中,以科内因素为主占1048份(71.83%),其中技术原因1017份(97.04%).科外因素411份,其中以责任原因为主,占270份(65.69%,X2=709.59,P<0.05).结论 影响病理报告及时性的因素有技术性和责任性两大类,科内以技术性为主,科外以责任性为主,应当分别运用流程改造、技术进步和加强责任心等措施加以改进.     

Evaluation of liquid and lyophilized preservatives for urine culture

Culture results of urine specimens transported conventionally (sterile cup) and in a commercial liquid or an investigational lyophilized preservative were compared in a hospital that experiences substantial delays in specimen transport to the laboratory (greater than 40% of specimens received after a delay of greater than or equal to 2 h). At the time of initial plating in the laboratory, 106 of 111 (95.5%) specimens that were positive (greater than or equal to 10(5) CFU of a single organism per ml in pure culture) after conventional transport were also positive in liquid preservative. After a 24-h holding period (cup refrigerated, preserved urine at room temperature), agreement was 91.4% (96 of 105). At the time of initial plating, agreement between results obtained by the conventional method and those obtained by using lyophilized preservative was 96.9% (63 of 65); after 24 h, agreement was 92.4% (61 of 67). Complete inhibition of growth of three Klebsiella pneumoniae isolates was observed in liquid preservative; however, urine processed in the lyophilized preservative did not show inhibition. The proportion of urine cultures showing no change in quantitative growth between the time of initial plating and repeat plating at 24 h was virtually identical for all three processing methods (83.6 +/- 0.9%). After the 24-h holding period, specimens processed in lyophilized preservative were less likely to show diminished quantitative growth than were specimens processed conventionally or in liquid preservative but were more likely to show an increase in growth of greater than or equal to 1 log. Nonetheless, the apparent lack of toxicity of lyophilized preservative may make it preferable to the currently available liquid preservative.     

Rapid detection of herpes simplex virus in clinical specimens by centrifugation and immunoperoxidase staining.

The effect of immunoperoxidase staining and centrifugation on the sensitivity and rapidity of herpes simplex virus detection in mink lung cell cultures was determined with 730 clinical specimens. In standard tube cultures, the use of immunoperoxidase staining resulted in detection of 31 (91%) of 34 positive cultures after overnight incubation, compared with 25 (74%) detected without the stain (P less than 0.05). The effect of centrifugation of specimens onto the monolayer followed by overnight incubation and immunoperoxidase staining was studied with 431 specimens. Of 107 positive specimens, 103 (96%) were detected by this method, compared with 91 (85%) detected in standard cell cultures observed for 5 days (P less than 0.02). Standard cell cultures that were examined after overnight incubation detected only 62 (58%) of the 107 positive specimens (P less than 0.001). Centrifugation of clinical specimens onto cell monolayers followed by overnight incubation and immunoperoxidase staining is more rapid and sensitive than are standard cell culture techniques for the laboratory diagnosis of herpes simplex virus infection.     

Filtration of platelet-poor plasma specimens yields platelet-free plasma. Application for batch analysis of beta-thromboglobulin and platelet factor 4

The authors investigated filtration of platelet-poor plasma through a 0.2-micron filter for the analysis of beta-thromboglobulin (beta-TG) and platelet factor 4 (PF4) in frozen specimens. Platelets were detected by electron micrograph in the platelet-poor plasma preparation without filtration. No platelets or platelet fragments were seen after filtration. Freezing the unfiltered specimens resulted in a significant increase in the beta-TG and PF4 concentrations, presumably because of release of these proteins from the platelet alpha-granules during freezing and thawing. There was no change in the PF4 levels and only a slight decrease in beta-TG levels after freezing of the filtered specimens. The filtered samples could be frozen for later batch analysis, resulting in considerable cost savings to the laboratory.     

Is the grading of breast carcinomas affected by a delay in fixation?

Summary The effect of delay in fixation on the modified Bloom and Richardson grade of eight breast carcinomas was investigated. Topologically shuffled samples of each tumour were immersed in fixative at times of 0.5, 2, 4, 6, 18 and 24 h after surgical removal. The grade of each tumour was assessed at delays of 0.5 and 6 h. The tubule formation and nuclear pleomorphism components of the grade showed no change with a delay in fixation of 6 h. The number of mitotic figures declined by a mean of 53% over the same period and this resulted in a decrease in the histological grade of one of the tumours. The implications of these findings for the handling of breast specimens in a diagnostic histopathological laboratory are discussed.Some of these data were presented at the 162nd meeting of the Pathological Society of Great Britain and Ireland at Cambridge, 3 January 1991. Published as an abstract in J Pathol (1991) 163:154A     

Impact of imipenem/cilastatin therapy on faecal flora

To evaluate the effects of parenteral imipenem/cilastatin therapy upon human faecal microflora, stool specimens obtained from ten patients before, during and after therapy were cultured quantitatively for aerobic and anaerobic microorganisms. The patients received 500 mg imipenem combined with 500 mg cilastatin every 6 h for 6–11 days. The antimicrobial therapy was associated with a small decrease in the numbers of enterobacteria, anaerobic cocci and bacteroides during treatment but afterward the microflora normalized in all patients. None of the patients was colonized with new imipenem-resistant bacteria, hadClostridium difficile or cytotoxin in the stools, or developed diarrhoea.     

Legionellosis

Although increasing attention is being given to Legionella pneumonia in Japan, reports of solitary onset of this disease are scant in Japan. The patient, from whom L. dumoffii was isolated, was a 59-year-old male with no underlying disease. He visited our hospital because of fever and cough, and was admitted to our department for X-ray findings consistent with pneumonia. After admission, pulmonary lesions spread rapidly, and based on the suspicion of Legionella pneumonia, drugs such as EM, RFP and MINO were used. However, the patient died on the 26th hospital day. L. dumoffii was isolated from specimens obtained by airway aspiration before death and specimens of lung abscess and airway discharge obtained during autopsy (7 specimens in total). In addition, the L. dumoffii antibody titer in the serum became elevated. This is the first case of L. dumoffii pneumonia reported in Japan. The other case was in an 81-year-old male with underlying disease. He was admitted urgently with suspected pneumonia but died on the following day. L. pneumophila serogroup 5 was isolated from autopsied lung tissue. Fatality is high for this disease, making early diagnosis and treatment with appropriate antibiotics essential. Physicians should bear in mind the possibility of this disease and request the necessary laboratory tests in suspected cases without delay.     

Wet mount microscopy reflects functional vaginal lactobacillary flora better than Gram stain

AIM: The status of vaginal lacto-bacillary flora, an indicator of possible genital infection and pregnancy complications, can be assessed on wet mount or Gram stained specimens. The former is quick, the latter more routine. The accuracy of the two preparative techniques to detect normal vaginal lacto-bacillary microflora was compared for 646 patients. The effect of delay in transport medium before Gram staining was also investigated. METHODS: Patients presented with infectious vaginitis or for a routine prenatal visit. After placement of a speculum, duplicate smears were taken from the upper vaginal vault and examined fresh or after Gram staining. Lacto-bacillary grades from both methods were compared with lactate concentration in vaginal rinses. In a subgroup of 238 patients, Gram staining was performed both on fresh smears and those that had been transported in Stuart's growth medium. RESULTS: Higher lacto-bacillary grades (more disrupted flora) were diagnosed 2.9 times more frequently on Gram stained specimens than on wet mounts (p < 0.0001), a difference even more pronounced after transport in Stuart's medium (relative risk, 4.2; p < 0.0001). Lacto-bacillary grades assessed on wet mounts correlated better with vaginal lactate concentration than those assessed on Gram stains. CONCLUSIONS: Easier recognition of lacto-bacillary morphotypes on wet mounts than on Gram stains might result from the loss of lactobacilli by the process of fixation or Gram staining. Wet mount microscopy of vaginal smears for assessment of lacto-bacillary grades, rather than Gram staining, is strongly recommended.     

Effect of centrifugation on herpes simplex virus isolation

The effects of high-speed centrifugation on the isolation of herpes simplex virus (HSV) were studied. Aliquots of laboratory or clinical specimens were inoculated into test tubes and flat-bottomed tubes containing HEp2 monolayers. Test tubes were incubated at 35 degrees C on roller drums (standard method), and flat-bottomed tubes were centrifuged at 15,000g at 35 degrees C for 1 hr, before being incubated at 35 degrees C without rolling (centrifuged method). Centrifugation of clinical and laboratory specimens of HSV type 1 and HSV type 2 produced significantly increased isolation rates compared with the standard method. When clinical and laboratory specimens were diluted, the centrifuged method was more sensitive at all dilutions. When 20 specimens were used for end-point titrations, the centrifuged method was 10 times more sensitive for 15 specimens and 100 times more sensitive for five specimens. There was no difference in the time taken for the appearance of cytopathic effect (CPE) between the standard and centrifuged methods.     

Differences in fecal microflora between patients with atopic dermatitis and healthy control subjects

BACKGROUND: The prevalence of allergic diseases, such as atopic dermatitis (AD), has been increasing. However, few investigations have been made of the intestinal microflora in Japanese patients with AD. OBJECTIVE: The purpose of this study was to determine the differences in microflora, fecal serum IgA concentrations, and skin IgA contents between patients with AD and healthy control subjects. METHODS: This trial was conducted as a case-control study using 30 minor patients with AD and age- and sex-matched healthy control subjects (n = 68). One week after a questionnaire was administered, fecal specimens and 24-hour skin secretion specimens were collected from all subjects. Fecal microflora, fecal IgA concentrations, and IgA contents on the skin surface were analyzed. RESULTS: The counts of Bifidobacterium (in log10 colony-forming units per gram) were significantly lower in patients with AD than in healthy control subjects (9.75 +/- 0.68 vs 10.10 +/- 0.50 log(10) colony-forming units/g, P <.05). In particular, percentages of Bifidobacterium were significantly lower in patients with severe skin symptoms than in those with mild skin symptoms (40% +/- 6% vs 19% +/- 6%, P <.05). In addition, the frequency of occurrence of Staphylococcus was significantly higher in patients with AD than in healthy control subjects (83% vs 59%, P <.05). There were no significant differences in fecal IgA content or IgA content on the skin between the 2 groups. CONCLUSION: Patients with AD had lower counts of Bifidobacterium than healthy control subjects, and the frequency of Staphylococcus was higher in patients with AD than in control subjects. Disorder of the intestinal microflora might play a role in the onset of AD and the aggravation of skin symptoms.     

Sampling of specimens for laboratory examinations

By the construction of hospital total system, marked developments, such as efficiency and labor-saving of work, have introduced to clinical practice and medical managements. Almost the same developments were also acquired by virtue of computerization in laboratory analysis. In our hospital, the Laboratory Information System (LIS) was connected in on-line mode with the Shared Hospital Information System (SHIS) and the Medical Data Administration System (MDAS). These systems resulted in the possession of the efficiency for laboratory process and in the diminution of various kinds of errors. However, the delay in the systematization was found on the problem in the preanalytical phase, especially on the collection of specimens. It is mainly because of multiformity of the circumstances reflected in every patient and in every ward, for example, variousness of clinicians' order for laboratory tests, many procedures and intricately scheduled timetable of the collection of blood. In order to obtain the more rapid and accurate test results, the construction of computerized system is also necessary for the collection and the transformation of blood. In this paper, problems concerning with the collection (sampling) of specimens were summarized.     

Effect of transport medium and transportation time on culture of Helicobacter pylori from gastric biopsy specimens.

To determine whether transportation time and use of a low budget transport medium (NaCl 0.9%) would influence culture of Helicobacter pylori from gastric biopsy specimens, upper gastrointestinal endoscopy was performed on 42 patients. The specimens were cultured and examined histologically, and H pylori antibodies were determined using an ELISA technique. Patients were regarded as H pylori positive when culture was positive or when histology or IgG anti-H pylori antibodies indicated H pylori infection. Rapid transportation of gastric biopsy specimens in NaCl 0.9%, at room temperature resulted in a high diagnostic yield (23 H pylori positive cultures in 26 patients with H pylori infection). A 24 hour delay in plating gastric biopsy specimens after transportation in NaCl 0.9%, at room temperature, did not seriously affect results (22 instead of 23 H pylori positive cultures). The culture results after transportation in Cairy-Blair medium were comparable with those after transportation in NaCl 0.9%, but because of availability, low cost, and ease of handling in the endoscopy department, NaCl 0.9% was preferred as transport medium. This study shows that for culture of H pylori from gastric biopsy specimens sterile saline is an adequate medium, and that transportation can be delayed for 24 hours without a significant loss of diagnostic yield.     

Ultrasound-enhanced latex immunoagglutination test (USELAT) for detection of capsular polysaccharide antigen of Neisseria meningitidis from CSF and plasma

AIMS: An ultrasonic instrument, the Immunosonic, was used to evaluate ultrasound-enhanced latex immunoagglutination testing (USELAT) for detection and serogroup determination of Neisseria meningitidis in clinical specimens. METHODS: Eighty-two CSF and EDTA blood specimens from patients with suspected meningococcal disease (MD) were tested by USELAT. Results were compared with routine laboratory tests for confirmation of MD and discrepant results were resolved by analysis of further laboratory and clinical data. RESULTS: Using the Wellcogen Bacterial Antigen Kit, USELAT was positive in 20 (24%) specimens. The resolved sensitivity of USELAT was 49% compared with 67% for PCR. There were no discrepancies between serogroups indicated by USELAT and serogroups confirmed by PCR or culture grouping. CONCLUSIONS: Although USELAT could be performed in laboratories without facilities for PCR testing, a specific ultrasonic instrument is necessary and some experience is required in interpreting results. The lower resolved sensitivity makes USELAT unsuitable as a stand-alone rapid test, and it added little value to standard laboratory culture with PCR testing.     

Stability of the Resident Microflora and the Bacteriocinogeny of Streptococcus mutans as Factors Affecting its Establishment in Specific Pathogen-Free Rats

The outcome of the experimental implantation of Streptococcus mutans strains in humans and animals is unpredictable, and neither success nor failure can be explained. It seems logical to assume that, apart from dietary and host factors, the characteristics of the S. mutans strain involved and those of the resident plaque microflora are important in colonization. For example, previous work in this laboratory suggested that bacteriocin production accounts at least in part for the establishment of an invading bacterium in a microbial ecosystem. In the present study, a complex specific pathogen-free Ny plaque ecosystem was obtained by the inoculation of specific pathogen-free rats with Actinomyces viscosus Ny1 and S. sanguis Ny101, and the establishment of S. mutans in such rats was then examined. It was found that bacteriocinogenic (bac(+)) strains generally colonized in much higher proportions than non-bacteriocinogenic (bac(-)) strains. Moreover, the longer the delay in introducing S. mutans, the poorer was its establishment. Shortly after inoculation of strains Ny1 and Ny101, there is probably a transient state in which microbial equilibrium has not been reached, but later the specific pathogen-free Ny system attains a stable climax community which more strongly resists invaders. The ability of a number of S. mutans strains to establish in such a climax community was then examined, and it was found that bac(+) strains generally established at a higher level than did bac(-) strains. In summary, it was concluded that, although the bac(+) state is an important property in the successful invasion of a plaque by S. mutans, the stability of the resident microflora is also an important factor.     

Evaluation of culture and the Gen-Probe PACE 2 assay for detection of Neisseria gonorrhoeae and Chlamydia trachomatis in endocervical specimens transported to a state health laboratory.

The Gen-Probe PACE 2 assay (Gen-Probe Inc., San Diego, Calif.) was compared with culture for the detection of Neisseria gonorrhoeae in endocervical specimens that were mailed to the laboratory. During mail transport, the specimens were exposed to extremes of hot and cold weather for several days before arriving in the laboratory. Specimens on culture plates deteriorated during transport, as evidenced by many dead gonococcus-like colonies. The manufacturer's recommendation for reporting PACE 2 assay-positive results was modified to create a suspicious category for samples with relative light units near the positive cutoff value. Of a total of 4,869 specimens tested, 30 were positive by both methods and 102 were positive only by the PACE 2 assay. These additional 102 positive specimens were likely to be true positives, as indicated by several lines of indirect evidence, including detailed probe competition analysis, patient history, and the lack of false-positive results in hand-delivered specimens. Although Gen-Probe Inc. indicates that specimens are stable for up to 7 days, N. gonorrhoeae was easily detectable by the PACE 2 assay after 1 month of incubation at room temperature in the PACE 2 transport buffer. We also compared the Gen-Probe PACE 2 assay for Chlamydia trachomatis with culture on endocervical specimens delivered by same-day courier. Of 398 endocervical specimens tested, the PACE 2 assay detected 19 of 20 culture-positive samples. Although the assay failed to detect one culture-positive sample, it was able to detect two very weak culture-suspicious samples. Finally, PACE 2 assays for N. gonorrhoeae and C. trachomatis performed on the same samples indicated that the coinfection rate was 40% for women attending five family planning clinics. We concluded that the Gen-Probe PACE 2 assay system should be considered for use in testing those specimens that are transported to the laboratory through the mail.