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Vojnosanitetski pregled
2015, vol. 72, br. 8, str. 670-676
jezik rada: engleski
vrsta rada: izvorni naučni članak
objavljeno: 30/08/2015
doi: 10.2298/vsp140612065N
Nov mikroskopski metod za analizu preparata bojenog po gramu u dijagnozi poremećaja vaginalne mikroflore
aVojnomedicinska akademija, Beograd + Univerzitet odbrane, Medicinski fakultet Vojnomedicinske akademije, Beograd
bDermatology Center Parmova & DCP-VENEX Centre, Ljubljana, Slovenia
cHuman Reproduction Center, Budva, Montenegro

e-adresa: dane_nenadic@yahoo.co.uk

Sažetak

Uvod/Cilj. Nugentov skor još uvek važi za zlatni standard u velikoj većini studija o proceni vaginalne flore i dijagnozi bakterijske vaginoze (BV). Cilj ovog rada bio je da se ustanovi da je mikroskopska analiza vaginalnog brisa preparata bojenog po Gramu na uvećanju '200, brza i jednostavna tehnika, lako primenljiva u svakodnevnoj praksi i da bolje odražava kompleksnost vaginalne flore od metodologije po Nugentu. Metode. Preparati bojeni po Gramu kod 394 asimptomatske trudnice (24-28 nedelja trudnoće) klasifikovani su na osnovu Nugentovih mikroskopskih kriterijuma (imerzija, uvećanje '1000). Slajdovi su ponovo analizirani pod imerzijom na uvećanju '200. Na osnovu polukvantitativne procene broja (celularnost) i odnosa štapićastih (duži-na > 1,5 μm) i malih bakterijskih formi (< 1.5 μm) sve ispitanice bile su podeljene u 6 grupa: hipercelularni (normal full - NF), srednje celularan (normal mid - NM), hipocelularan (normal empty - NE), 'bacterial vaginosis full' (BVF), 'bacterial vaginosis mid' (BVM), i 'bacterial vaginosis empty' (BVE). Takođe, identifikovane su gljivice, koke, bifido i lepto bakterijske forme kao i polimorfonuklearni leukociti (PMN). Rezultati. Na osnovu Nugentovih kriterijuma, BV nađena je kod 78 ispitivanih trudnica, intermedijerni nalaz kod 63, i gljivice kod 48. Na osnovu naših kriterijuma BV je potvrđena kod 88 ispitanica (37 BVF, 24 BVM, i 27 BVN). Generalno, oba pristupa pokazala su visoku podudarnost u dijagnozi BV (Linov koeficijent podudarnosti korelacije = 0,9852). Kod 40% žena pronađena je mešovita flora: gljivice kod 126 (32%), koke kod 145 (37%), bifido forme kod 32 (8%) i lepto forme kod 20 (5%). Skoro polovina žena sa BV imala je, takođe, gljivice (39/88). Povećan broj PMN je nađen kod 102 (33%) trudnica sa normalnim nalazom i kod 36 (41%) trudnica sa BV. Zaključak. Nova metodologija jednostavnija je za primenu i mnogo bolje odražava raznolikost vaginalne mikroflore. Ovakav pristup možda bi bio korisniji molekularnim biolozima i njihovim pokušajima da na osnovu kvantitativne lančane reakcije polimeraze (polymerase chain reaction - PCR) dođu do molekularne 'formule' za dijagnostiku BV.

Ključne reči

Reference

Aagaard, K., Riehle, K., Ma, J., i dr. (2012) A Metagenomic Approach to Characterization of the Vaginal Microbiome Signature in Pregnancy. PloS One, 7(6): 36466
Amsel, R., Totten, P.A., Spiegel, C.A., Chen, K.C., Eschenbach, D., Holmes, K.K. (1983) Nonspecific vaginitis: Diagnostic criteria and microbial and epidemiologic associations. Am J Med, 74(1): 14-22
Biagi, E., Vitali, B., Pugliese, C., Candela, M., Donders, G.G.G., Brigidi, P. (2008) Quantitative variations in the vaginal bacterial population associated with asymptomatic infections: a real-time polymerase chain reaction study. European Journal of Clinical Microbiology & Infectious Diseases, 28(3): 281-285
Bradshaw, C. S., Tabrizi, S. N., Fairley, C. K., Morton, A. N., Rudland, E., Garland, S. M. (2006) The Association of Atopobium vaginae and Gardnerella vaginalis with Bacterial Vaginosis and Recurrence after Oral Metronidazole Therapy. Journal of Infectious Diseases, 194(6): 828-836
Brotman, R.M., Ravel, J., Cone, R.A., Zenilman, J.M. (2010) Rapid fluctuation of the vaginal microbiota measured by Gram stain analysis. Sex Transm Infect, 86(4): 297-302
Cartwright, C.P., Lembke, B.D., Ramachandran, K., Body, B.A., Nye, M.B., Rivers, C.A., Schwebke, J.R. (2012) Development and validation of a semiquantitative, multitarget PCR assay for diagnosis of bacterial vaginosis. Journal of Clinical Microbiology, 50(7): 2321-9
Cartwright, C.P., Lembke, B.D., Ramachandran, K., Body, B.A., Nye, M.B., Rivers, C.A., Schwebke, J.R. (2013) Comparison of Nucleic Acid Amplification Assays with BD Affirm VPIII for Diagnosis of Vaginitis in Symptomatic Women. Journal of Clinical Microbiology, 51(11): 3694-3699
de Backer, E., Verhelst, R., Verstraelen, H., Alqumber, M.A., Burton, J.P., Tagg, J.R., i dr. (2007) Quantitative determination by real-time PCR of four vaginal Lactobacillus species. Gardnerella vaginalis and Atopobium vaginae indicates an inverse relationship between L. gasseri and L. iners. BMC Microbiol, 19(7): 115
Forsum, U., Jakobsson, T., Larsson, P.G., Schmidt, H., Beverly, A., Bjørnerem, A., Carlsson, B., Csango, P., Donders, G., Hay, P., Ison, C., Keane, F., McDONALD, H., Moi, H., Platz-Christensen, J.-J., Schwebke, J. (2002) An international study of the interobserver variation between interpretations of vaginal smear criteria of bacterial vaginosis. APMIS, 110(11): 811-818
Fredricks, D.N., Fiedler, T.L., Thomas, K.K., Mitchell, C.M., Marrazzo, J.M. (2009) Changes in Vaginal Bacterial Concentrations with Intravaginal Metronidazole Therapy for Bacterial Vaginosis as Assessed by Quantitative PCR. Journal of Clinical Microbiology, 47(3): 721-726
Fredricks, D.N., Fiedler, T.L., Marrazzo, J.M. (2005) Molecular identification of bacteria associated with bacterial vaginosis. N Engl J Med, 353(18): 1899-911
Ison, C.A., Hay, P.E. (2002) Validation of a simplified grading of Gram stained vaginal smears for use in genitourinary medicine clinics. Sex Transm Infect, 78(6): 413-5
Lamont, R.F., Sobel, J.D., Akins, R.A., Hassan, S.S., Chaiworapongsa, T., Kusanovic, J.P., Romero, R. (2011) The vaginal microbiome: new information about genital tract flora using molecular based techniques. BJOG, 118(5): 533-49
Larsson, P. (2004) Diagnosis of bacterial vaginosis: need for validation of microscopic image area used for scoring bacterial morphotypes. Sexually Transmitted Infections, 80(1): 63-67
Libman, M.D., Kramer, M., Platt, R. (2006) Comparison of Gram and Kopeloff stains in the diagnosis of bacterial vaginosis in pregnancy. Diagnostic Microbiology and Infectious Disease, 54(3): 197-201
Lin, L.I. (1989) A Concordance Correlation Coefficient to Evaluate Reproducibility. Biometrics, 45(1): 255
Ling, X.Z., Kong, M.J., Liu, F., Zhu, B.H., Chen, Y.X., Wang, Z.Y., et al. (2010) Molecular analysis of the diversity of vaginal microbiota associated with bacterial vaginosis. BMC Genomics, 11(1): 488
Mcdonald, H.M., Brocklehurst, P., Gordon, A. (2007) Antibiotics for treatin g bacterial vaginosis in pregnancy. Cochrane Database Syst Rev., CD00026
Menard, J.P., Mazouni, C., Fenollar, F., Raoult, D., Boubli, L., Bretelle, F. (2010) Diagnostic accuracy of quantitative real-time PCR assay versus clinical and Gram stain identification of bacterial vaginosis. European Journal of Clinical Microbiology & Infectious Diseases, 29(12): 1547-1552
Menard, J.-P., Fenollar, F., Raoult, D., Boubli, L., Bretelle, F. (2011) Self-collected vaginal swabs for the quantitative real-time polymerase chain reaction assay of Atopobium vaginae and Gardnerella vaginalis and the diagnosis of bacterial vaginosis. European Journal of Clinical Microbiology & Infectious Diseases, 31(4): 513-518
Menard, J., Fenollar, F., Henry, M., Bretelle, F., Raoult, D. (2008) Molecular Quantification of Gardnerella vaginalis and Atopobium vaginae Loads to Predict Bacterial Vaginosis. Clinical Infectious Diseases, 47(1): 33-43
Nugent, R.P., Krohn, M.A., Hillier, S.L. (1991) Reliability of diagnosing bacterial vaginosis is improved by a standardized method of gram stain interpretation. J Clin Microbiol, 29(2): 297-301
Pereira, L., Culhane, J., Mccollum, K., Agnew, K., Nyirjesy, P. (2005) Variation in microbiologic profiles among pregnant women with bacterial vaginosis. Am J Obstet Gynecol, 193(3): 746-751
Ravel, J., Gajer, P., Abdo, Z., Schneider, G.M., Koenig, S.S., Mcculle, S.L., i dr. (2011) Vaginal microbiome of reproductive-age women. Proc Natl Acad Sci USA, 108(1): 4680-4687, Suppl
Shipitsyna, E., Roos, A., Datcu, R., Hallén, A., Fredlund, H., Jensen, J.S., Engstrand, L., Unemo, M. (2013) Composition of the Vaginal Microbiota in Women of Reproductive Age – Sensitive and Specific Molecular Diagnosis of Bacterial Vaginosis Is Possible?. PLoS ONE, 8(4): e60670
Sobel, J.D., Subramanian, C., Foxman, B., Fairfax, M., Gygax, S.E. (2013) Mixed Vaginitis—More Than Coinfection and With Therapeutic Implications. Current Infectious Disease Reports, 15(2): 104-108
Srinivasan, S., Morgan, M.T., Liu, C., Matsen, F.A., Hoffman, N.G., Fiedler, T.L., i dr. (2013) More than meets the eye: associations of vaginal bacteria with gram stain morphotypes using molecular phylogenetic analysis. PLoS One, 8(10): 78633
Srinivasan, S., Hoffman, N.G., Morgan, M.T., Matsen, F.A., Fiedler, T.L., Hall, R.W., i dr. (2012) Bacterial Communities in Women with Bacterial Vaginosis: High Resolution Phylogenetic Analyses Reveal Relationships of Microbiota to Clinical Criteria. PloS One, 7(6): 37818
Thies, F.L., König, W., König, B. (2007) Rapid characterization of the normal and disturbed vaginal microbiota by application of 16S rRNA gene terminal RFLP fingerprinting. J Med Microbiol, 56(Pt 6): 755-61
Tohill, B.C., Heilig, C.M., Klein, R.S., Rompalo, A., Cu-Uvin, S., Brown, W., Duerr, A. (2004) Vaginal flora morphotypic profiles and assessment of bacterial vaginosis in women at risk for HIV infection. Infect Dis Obstet Gynecol, 12(3-4): 121-6
Verhelst, R., Cools, P., Lopes dos Santos Santiago G,, Temmerman, M., Veneechoutte, M. (2011) Garnderella. u: Liu D. [ur.] Molecular detection of human bacterial pathogens, Boca Raton: Press Taylor & Fracis Group, str. 81-95
Verhelst, R., Verstraelen, H., Claeys, G., Verschraegen, G., van Simaey, L., de Ganck, C., et al. (2005) Comparison between Gram stain and culture for the characterization of vaginal microflora: Definition of a distinct grade that resembles grade I microflora and revised categorization of grade Imicroflora. BMC Microbiology, 5(1): 61
Zhou, X., Brotman, R.M., Gajer, P., Abdo, Z., Schüette, U., Ma, S., i dr. (2010) Recent advances in understanding the microbiology of the female reproductive tract and the causes of premature birth. Infect Dis Obstet Gynecol, 2010: 737425
Zhou, X., Bent, S.J., Schneider, M.G., Davis, C.C., Islam, M.R., Forney, L.J. (2004) Characterization of vaginal microbial communities in adult healthy women using cultivation-independent methods. Microbiology, 150(Pt 8): 2565-73
Zhou, X., Brown, C.J., Abdo, Z., Davis, C.C., Hansmann, M.A., Joyce, P., Foster, J.A., Forney, L.J. (2007) Differences in the composition of vaginal microbial communities found in healthy Caucasian and black women. ISME J, 1(2): 121-33